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The Practical Manual of Histology, Second Edition, is designed for first-year MBBS students in India, aligning with the competency-based curriculum. It provides a comprehensive guide to identifying and describing human tissues under a microscope, featuring hand-drawn diagrams and specific learning objectives. The manual aims to facilitate easier learning and preparation for histology assessments, including the NEXT exam.

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0% found this document useful (0 votes)

197 views323 pages

Hina Sharma - Practical Manual of Histology, 2e (2022) - 2

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Practical Manual of

Histology Second Edition

As per CBME Guidelines | Competency Based Undergraduate

Curriculum for the Indian Medical Graduate

Name ............................................................................................................................................................
Year ................................................................................Roll no. ...........................................................
Name of the College .............................................................................................................................
Teacher’s Signature ......................................................................................................................................

This is to certify that Mr/Ms 1st year MBBS student of batch has
attained the knowledge and skill to identify all types of epithelia under the microscope and describe the various
types that correlate to its function.

Signature and seal of the Head Signature and seal of the Dean
Department of Anatomy

This is to certify that Mr/Ms 1st year MBBS student of batch has
attained the knowledge and skill to identify, draw and label a slide of trachea and lung.

Signature and seal of the Head Signature and seal of the Dean
Department of Anatomy
Practical Manual of
Histology Second Edition

As per CBME Guidelines | Competency Based Undergraduate

Curriculum for the Indian Medical Graduate

Hina Sharma MBBS, MD (Anatomy)

Associate Professor
Department of Anatomy
Geetanjali Medical College and Hospital
Udaipur, Rajasthan, India

CBS Publishers & Distributors Pvt Ltd

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Disclaimer
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information and knowledge. The authors have tried their best in giving information available to them while
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material, yet it is quite possible some errors might have been left uncorrected. The publisher, the printer and
the authors will not be held responsible for any inadvertent errors, omissions or inaccuracies.

eISBN: xxxx
Copyright © Authors and Publisher

Second eBook Edition: 2022

All rights reserved. No part of this eBook may be reproduced or transmitted in any form or by any means,
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without permission, in writing, from the authors and the publisher.

Published by Satish Kumar Jain and produced by Varun Jain for

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Foreword

I t gives me immense pleasure to introduce the Practical Manual of Histology as per the latest MCI syllabus.
The author of this workbook has written it with the purpose of dealing with the practical aspects of identifying
and describing the normal microanatomy of human tissues.
It has a good consortium of specific learning objectives, think and answer, three points of identification and
hand-drawn diagrams. The author has addressed the problems encountered by the learners and penned this
workbook in a way which will engender faster understanding of the subject.
I hope the students will take full advantage of the crisp dose of knowledge and diagrams provided in this
workbook. At a first glance it looks very promising indeed and I wish the author success in her intent of making
teaching and learning of histology a fruitful and rewarding experience.

Prof FS Mehta MS (General Surgery)

Dean, Geetanjali Medical College and Hospital
Udaipur, Rajasthan
former Superintendent
Rabindra Nath Tagore Medical College and Hospital
Udaipur, Rajasthan, India
Preface to the Second Edition

I am grateful to the Almighty for giving me the opportunity to present the second edition of Practical Manual of
Histology as per the new guidelines of competency based undergraduate curriculum for the Indian medical
graduate.
I am glad for the immense heartening response from my colleagues, seniors, juniors, students, and my teachers.
It has given me the strength to improvise on it and come with the second edition. I hope and look forward to
similar and more acceptance from the stakeholders.
The features improvised in the second edition are as follows:
1. The color of the diagrams has been made lighter, as seen in H&E staining.
2. Each diagram has been named.
3. The diagrams which have been revised are:
a. Small diagrams of cells, collagen fibres, elastic fibres, serous acini and mucous acini and stratified cuboidal
epithelium.
b. Hyaline cartilage
c. Skeletal muscle
d. Thyroid gland
e. Urinary bladder
f. Fallopian tube
4. The extra blank pages have been removed.
5. SDL (Self-directed Learning) pages have been removed.
6. The text and diagram pages have been rearranged to lie next to each other, so that the learner does not have
to turn the page to see the features mentioned in the text.
It is my humble effort to make learning of histology easy for the overburdened medical graduate learners. This
manual shall be useful in attempting histology questions in the proposed NEXT exam apart from being a complete
book in itself for the 1st year of preclinical subjects.

Hina Sharma
Preface to the First Edition

D uring the years of teaching histology I have experienced the difficulties encountered by the learners. There is
a vast choice of literature for the students to pick from. The normal tendency to pick the easiest available
means leads to notes borrowed from seniors. It leads to the problem seen in the whispering game—the end
product is far from correct.
Another practical problem encountered is that the various textbooks offer various stains, whereas the stain
usually used and taught is haematoxylin and eosin (H&E). As such the learner is faced with the dilemma of
which diagram to draw.
To address these issues the idea of promoting easy learning by providing hand-drawn diagrams of tissues
stained with H&E stain was conceived. The diagrams are self-explanatory and help the learner identify the given
tissue.
The highlight of this workbook is that it addresses all the latest competencies of histology given in the new
curriculum.
Each slide has been given specific learning objectives. So at the very outset the learner knows what all he has to
specifically look for in the given slide.
There are think and answer questions given at places to encourage the learner to use his/her grey cells because
the mind sees what the brain knows. So once the learner begins with thinking he knows what to look for.
There is space provided for writing three points of identification of the slide. These will prove invaluable for
last moment revision prior to assessments.
SDL pages have been provided for notes or diagrams. Sufficient space has been given to draw the diagrams
with exemplary diagram above on the same page, thus removing the tedious process of turning pages to copy
while drawing. If smaller diagram is to be drawn, then a smaller circle may be drawn within the larger one.
This workbook is ideal for learning histology in various streams related to health education and is sufficient
for preparation of viva voce.

Hina Sharma
Acknowledgements

W riting a book is by far harder than I first thought. Nonetheless it is very rewarding. I thank the Almighty
for giving me the opportunity to write this book.
I am eternally grateful to the management of Geetanjali Medical College for encouraging academic excellence
and supporting in adopting newer methods of teaching. I am specially grateful to the Dean, Prof FS Mehta, for
initiating the idea of writing a workbook, without which I would probably have not even thought of it. He has
been very supportive and inspired me to start this work.
Very special thanks to Prof Manjinder Kaur, Head, Department of Physiology, Geetanjali Medical College, for
giving me the much required momentum to start writing this workbook. Being the academic officer, she has
always been a source of stimulation for academic perfection.
I cannot thank my family, husband Dr DK Sharma, daughter Dr Ila Sharma and son Dr Kartikeya, enough for
bearing with me and not being impatient when I could not do some of the chores due to preoccupation. Their
constant support was the only reason that this workbook could be completed on time.
My teacher and guide Prof LK Jain has always been a source of inspiration. The knowledge imparted by him is
sound, complete and dependable. I thank him for his blessings.
I am grateful to Prof Nikha Bharadwaj for her invaluable support and suggestions.
My colleagues Dr Brijesh, Dr Charu, Dr Meghna and Dr Pritesh have provided an amicable environment and
encouraged and appreciated my work. I am very grateful for their reinforcement.
My initial teaching years were very enriching and I thank my then colleagues Prof Ghanshyam Gupta,
Prof Seema Prakash, Dr Parveen Ojha and Dr Pooja Dhabai for providing an academically inspiring environment.
I thank Prof GC Agarwal for helping me enhance my knowledge of histology during initial teaching years. He
encouraged me to avoid taking a casual approach towards teaching histology.
I wholeheartedly thank Mrs Jyotsana, Mr Biju, Mr Bhagwati, Mr Vinod and Mr Jagdish for their support.
The students have always been a source of inspiration. I thank them for putting their faith in me and giving me
the much required compliments.
I am grateful to all my friends for encouraging me to complete this workbook.
I thank the publisher, and the others involved for printing this workbook in such a short notice.

Hina Sharma
Contents
Foreword by Prof FS Mehta v
Preface to the Second Edition vii
Preface to the First Edition ix
Index of Competencies for Histology xix
Abbreviations xx

No. Practical Page no. Teacher’s signature Remarks

1 Light Microscope 1

2 Epithelium Histology 4

3 Classification of Epithelium 4

4 Simple Squamous Epithelium 8

5 Simple Cuboidal Epithelium 10

6 Simple Columnar Epithelium 12

7 Simple Ciliated Columnar Epithelium 14

8 Simple Columnar Epithelium with Brush/Striated Border 16

9 Stratified Squamous Epithelium (Non-keratinized) 18

10 Stratified Squamous Epithelium (Keratinized) 20

11 Stratified Cuboidal Epithelium 22

12 Stratified Columnar Epithelium 24

13 Transitional Epithelium (Unstretched Urinary Bladder) 26

14 Transitional Epithelium (Stretched Urinary Bladder) 28

15 Pseudostratified Ciliated Columnar Epithelium (Trachea) 30

16 Olfactory Epithelium 33

17 Ultrastructure of Epithelium 36

18 Connective Tissue Histology 40

19 Loose Areolar Tissue 42

20 Dense Regular Connective Tissue (Tendon LS) 44

21 Dense Regular Connective Tissue (Tendon TS) 46

22 Ultrastructure of Connective Tissue 48

23 Cartilage 50
xiv Practical Manual of Histology

No. Practical Page no. Teacher’s signature Remarks

24 Hyaline Cartilage 52

25 Elastic Cartilage 54

26 Fibrocartilage 56

27 Bone 59

28 Compact Bone (TS) 60

29 Compact Bone (LS) 62

30 Cancellous Bone (Spongy Bone) TS 64

31 Muscular Tissue 67

32 Skeletal Muscle (LS) 68

33 Skeletal Muscle (TS) 70

34 Cardiac Muscle (LS) 72

35 Smooth Muscle (LS) 74

36 Smooth Muscle (TS) 76

37 Muscle (Ultrastructure) 78

38 Nervous Tissue 80

39 Peripheral Nerve (TS) 82

40 Sensory/Spinal/Dorsal Root Ganglion 84

41 Autonomic/Sympathetic/Parasympathetic Ganglion 86

42 Exocrine (Salivary) Glands Histology 88

43 Sublingual Salivary Gland (Mucous Gland) 90

44 Parotid Salivary Gland (Serous Gland) 92

45 Submandibular Salivary Gland (Mixed Gland) 94

46 Blood Vessels Histology 96

47 Elastic Artery/Large Artery 98

48 Muscular Artery/Medium-sized Artery 100

49 Lymphoid Tissue 102

50 Lymph Node 103

51 Spleen 107

52 Thymus 111
Contents xv

No. Practical Page no. Teacher’s signature Remarks

53 Palatine Tonsil 115

54 Integumentary System 119

55 Thin Skin 120

56 Thick Skin 122

57 Respiratory System 125

58 Trachea 126

59 Lung 129

60 Interalveolar Septum 133

61 Epiglottis 135

62 Gastrointestinal System 138

63 Tongue 139

64 Oesophagus 143

65 TS through Lower One-third of Oesophagus 144

66 TS through Upper One-third of Oesophagus 146

67 TS through Middle One-third of Oesophagus 148

68 Cardioesophageal Junction 151

69 Stomach 155

70 Fundus/Body of Stomach (TS) 156

71 TS through Pylorus of Stomach 159

72 Intestines 163

73 Duodenum 164

74 Jejunum 166

75 Ileum 168

76 Large Intestine 170

77 Vermiform Appendix 171

78 TS through Vermiform Appendix 172

79 Colon 174

80 Hepatobiliary System 177

81 Liver 178
xvi Practical Manual of Histology

No. Practical Page no. Teacher’s signature Remarks

82 Pancreas 181

83 Gall Bladder 185

84 Urinary System 188

85 Kidney 190

86 Ureter 193

87 Urinary Bladder 197

88 Reproductive System 201

89 Male Reproductive System 201

90 Testis 202

91 Epididymis 205

92 Vas Deferens/Ductus Deferens 209

93 Vas Deferens 210

94 Prostate 213

95 Penis 217

96 Female Reproductive System 221

97 Ovary 222

98 Uterus 225

99 Uterus (Proliferative Phase) 226

100 Uterus (Secretory Phase) 228

101 Cervix 231

102 Fallopian Tube/Uterine Tube 235

103 Placenta 239

104 Umbilical Cord 242

105 Corpus Luteum 245

106 Breast 249

107 Mammary Gland (Resting) 250

108 Mammary Gland of Pregnancy 252

109 Mammary Gland of Lactation 254

Contents xvii

No. Practical Page no. Teacher’s signature Remarks

110 Endocrine Glands 257

111 Pituitary Gland 257

112 Thyroid Gland 260

113 Parathyroid Gland 262

114 Pineal Gland 264

115 Suprarenal Gland 267

116 Central Nervous System 270

117 Spinal Cord 270

118 Cerebrum 272

119 Cerebellum 274

120 Special Senses 277

121 Cornea 277

122 Sclero-corneal Junction 281

123 Retina 285

124 Optic Nerve 288

125 Eyelid 291

126 Lip 294

127 Cochlea and Organ of Corti 297

Guide to Students for Staining Characteristics of Histology Tissues 300

Feedback form 301

SUGGESTED ASSESSMENT CRITERIA

• Complete file with good and accurate diagrams 5 marks
• Complete file but a few (less than 10%) incorrect diagrams 4 marks
• Complete file but mostly (more than 50%) incorrect diagrams 3 marks
• Incomplete file (more than 75% complete) 2 marks
• Incomplete file (less than 75% complete) 1 mark
• No diagrams drawn (less than 25% diagrams drawn) Zero
INDEX OF COMPETENCIES FOR HISTOLOGY

Number Competencies addressed Page nos

AN9.2 Breast: Describe the location, extent, deep relations, structure, age changes, 249–255
blood supply, lymphatic drainage, microanatomy and applied anatomy of breast
AN25.1 Identify, draw and label a slide of trachea and lung 125–131
AN43.2 Identify, describe and draw the microanatomy of pituitary gland, thyroid, 257–263, 139–142, 88–95,
parathyroid gland, tongue, salivary glands, tonsil, epiglottis, cornea, retina 115–117, 136–137, 178–187
AN43.3 Identify, describe and draw microanatomy of olfactory epithelium, eyelid, 33–35, 291–295, 288–289,
lip, sclera-corneal junction, optic nerve, cochlea-organ of Corti, pineal gland 297–299, 264–265
AN52.1 Describe and identify the microanatomical features of gastro-intestinal system: 143–187, 268–269
Oesophagus, fundus of stomach, pylorus of stomach, duodenum, jejunum,
ileum, large intestine, appendix, liver gall bladder, pancreas and suprarenal gland
AN52.2 Describe and identify the microanatomical features of: 188–248
Urinary system: Kidney, ureter and urinary bladder
Male reproductive system: Testis, epididymis, vas deferens, prostate and penis
Female reproductive system: Ovary, uterus, uterine tube, cervix,
placenta and umbilical cord
AN52.3 Describe and identify the microanatomical features of cardioesophageal junction, 151–153, 245–247
corpus luteum
AN64.1 Describe and identify the microanatomical features of spinal cord, 270–275
cerebellum and cerebrum
AN65.1 Identify epithelium under the microscope and describe the various types 4–35
that correlate to its function
AN65.2 Describe the ultrastructure of epithelium 36–39
AN66.1 Describe and identify various types of connective tissue with functional correlation 40–47
AN66.2 Describe the ultrastructure of connective tissue 48–49
AN67.1 Describe and identify various types of muscle under the microscope 67–79
AN67.2 Classify muscle and describe the structure–function correlation of the same 78–79
AN67.3 Describe the ultrastructure of muscular tissue 78–79
AN68.1 Describe and identify multipolar and unipolar neuron, ganglia, peripheral nerve 80–83
AN68.2 Describe the structure–function correlation of neuron 80–81
AN68.3 Describe the ultrastructure of nervous tissue 80–81
AN69.1 Identify elastic and muscular blood vessels, capillaries under the microscope 96–101
AN69.2 Describe the various types and structure–function correlation of blood vessels 96
AN69.3 Describe the ultrastructure of blood vessels 97
AN70.1 Identify exocrine gland under the microscope and distinguish between serous, 88–95
mucous and mixed acini
AN70.2 Identify the lymphoid tissue under the microscope and describe microanatomy 102–117
of lymph node, spleen, thymus, tonsil and correlate the structure with function
AN71.1 Identify bone under the microscope; classify various types and describe 59–65
structure–function correlation of the same
AN71.2 Identify cartilage under the microscope and describe various types and 50–57
structure–function correlation of the same
AN72.1 Identify the skin and its appendages under the microscope and correlate the 119–123
structure with function

Note: Competency numbers are as provided in the new curriculum.

ABBREVIATIONS

1. SLO: Specific learning objectives

2. H&E: Haematoxylin and eosin
3. LS: Longitudinal section
4. ECM: Extracellular matrix
5. BM: Bone marrow
6. LN: Lymph node
7. WBC: White blood cells
8. RBCs: Red blood cells
9. RER: Rough endoplasmic reticulum
10. SER: Smooth endoplasmic reticulum
Practical Manual of Histology 1

Date:

LIGHT MICROSCOPE

LABEL THE PARTS OF THE MICROSCOPE

1
2 Practical Manual of Histology

Date:

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify all parts of the microscope.
2. Tell the functions of all the parts.
3. Attain the skill to focus a given slide at low and high power.

ACTIVITY
1. Focus the given slide in low power (10X)
2. Focus the given slide in high power (40X)

Viva Questions
1. Name the various types of microscopes

2. Name the parts of a light microscope used in your lab.

3. What are the magnifications available in the given light microscope? Which ones are commonly used?

4. How do you focus a slide on a light microscope?

5. What precautions are taken while focussing a slide in the light microscope?
Practical Manual of Histology 3

Date:

6. Which magnification is used first while focussing a slide?

SHOW HOW
1. Show how you adjust the lens for coarse focussing.
2. Show how you adjust the lens for fine focussing.
3. Which side of the slide do you keep upwards?

Note
Whenever you focus and try to identify a slide, always see the whole tissue by moving the slide on the stage of
the microscope. Do not comment on the slide by looking at just one field.
4 Practical Manual of Histology

Date:

EPITHELIUM HISTOLOGY

Number Competency Domain Level Core Teaching/ Assessment Number required

(Y/N) learning method method to certify
AN 65.1 Identify epithelium under K/S P Y Lecture, practical Written/skill 1
the microscope and describe assessment
the various types that
correlate to its function

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Classify different types of epithelia.

CLASSIFICATION OF EPITHELIUM
Answer the Following
Simple Epithelium (Single Layer of Cells)
Flat cells (squamous)
1. For example

2. What is the shape of the nucleus in such cells?

3. What is endothelium?

4. What is mesothelium?

Cells with equal height and width (cuboidal)

5. For example
Practical Manual of Histology 5

Date:

6. What is the shape of the nucleus in such cells?

7. Where does the basement membrane lie?

Cells with height more than width (columnar)

8. For example

9. What is the shape of the nucleus in such cells?

10. Where does the basement membrane lie?

Stratified Epithelium (Multilayered Cells)

Stratified squamous epithelium
11. Shape of deeper layer of cells is:

12. What is the shape of the nucleus in such cells?

13. Shape of surface layer of cells is:

6 Practical Manual of Histology

Date:

14. For example

15. Where does the basement membrane lie?

Stratified cuboidal epithelium

16. Shape of deeper layer of cells is:

17. What is the shape of the nucleus in such cells?

18. Shape of surface layer of cells is:

19. For example

20. Where does the basement membrane lie?

Stratified columnar epithelium

21. Shape of deeper layer of cells is:

22. What is the shape of the nucleus in such cells?

Practical Manual of Histology 7

Date:

23. Shape of surface layer of cells is:

24. For example

25. Where does the basement membrane lie?

Transitional epithelium

26. Shape of deeper layer of cells is:

27. What is the shape of the nucleus in such cells?

28. Shape of surface layer of cells is:

Three points of identification

Practical Manual of Histology 29

Date:

Transitional epithelium (stretched urinary bladder)

30 Practical Manual of Histology

Date:

PSEUDOSTRATIFIED CILIATED COLUMNAR EPITHELIUM (TRACHEA)

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Describe the shape of cells and nuclei in pseudostratified columnar epithelium.
2. Give examples of tissues with pseudostratified ciliated columnar epithelium.
3. Describe what cilia are.
4. Describe the function of this epithelium.
5. Draw a diagram showing microanatomy of pseudostratified ciliated columnar epithelium.

Distribution and function

Three points of identification

Practical Manual of Histology 31

Date:

Pseudostratified ciliated columnar epithelium (trachea)

32 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 33

Date:

OLFACTORY EPITHELIUM

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 43.2 Identify, describe and draw K/S SH Y Lecture, practical Written/skill
the microanatomy of assessment
pituitary gland, thyroid,
parathyroid gland, tongue,
salivary glands, tonsil,
epiglottis, cornea, retina

• Olfactory epithelium is present on the superior nasal concha and adjacent nasal septum near the roof of nasal
cavity.
• The epithelium is pseudostratified. The superficial cytoplasm appears clear and the nuclear layer shows many
rows of nuclei.
• Nuclei of olfactory cells are round. The superficial parts of these cells have non-motile olfactory cilia.
• Other cells of epithelium are sustentacular cells and basal cells.
• Lamina propria lies deep to epithelium. It has blood capillaries, lymphatics, nerves, and serous glands called
Bowman’s glands.
34 Practical Manual of Histology

Date:

OLFACTORY EPITHELIUM

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify olfactory epithelium under the light microscope.
2. Identify pseudostratified epithelium of olfactory mucosa.
3. Identify olfactory neuron.
4. Identify sustentacular and basal cells.
5. Identify Bowman’s glands.
6. Draw a diagram of microanatomy of olfactory epithelium.

Distribution and function

Three points of identification

Practical Manual of Histology 35

Date:

Olfactory epithelium
36 Practical Manual of Histology

Date:

ULTRASTRUCTURE OF EPITHELIUM

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 65.2 Describe the ultrastructure K KH N Lecture, practical Written
of epithelium

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Describe ultrastructure of basement membrane.
2. Describe ultrastructure of cilia.
3. Describe ultrastructure of microvilli.
4. Describe ultrastructure of stereocilia.

I. ULTRASTRUCTURE OF BASEMENT MEMBRANE

1. Name the layers of basement membrane as seen under the electron microscope.

2. Write two functions of stereocilia.

3. Where are stereocilia found?

40 Practical Manual of Histology

Date:

CONNECTIVE TISSUE HISTOLOGY

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 66.1 Describe and identify various K/S SH Y Lecture, practical Written/skill
types of connective tissue with assessment
functional correlation

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Classify connective tissue.
2. Name the components of connective tissue.
3. Name the three types of fibres in connective tissue.
4. Name the types of proper connective tissues with examples.

Classification of Connective Tissue

1. Ordinary
2. Loose areolar connective tissue, e.g. subperitoneal tissue, endomysium, lamina propria.
3. Dense collagenous
a. Dense regular connective tissue, e.g. tendon, ligaments, aponeurosis
b. Dense irregular connective tissue, e.g. reticular layer of dermis
4. Connective tissue with special properties
a. Adipose, e.g. subcutaneous adipose tissue
b. Mucoid, e.g. Wharton’s jelly
c. Reticular, e.g. stroma of lymphoid and myeloid (bone marrow) organs
d. Elastic, e.g. blood vessels, ligamentum nuchae
e. Scleral
i. Cartilage
ii. Bone
5. Lymphoid
6. Haemopoietic
Practical Manual of Histology 41

Date:

COMPONENTS OF CONNECTIVE TISSUE PROPER

• Name the cells found in connective tissue:

At the end of the session the student should be able to:
1. Describe the ultrastructure of ground substance of connective tissue.
2. Describe the ultrastructure of collagen fibres and their types.
3. Describe the ultrastructure of elastic fibres.
4. Describe the ultrastructure of reticular fibres.

GROUND SUBSTANCE
• Ground substance is made up of glycosaminoglycans (mucopolysaccharides), proteoglycans and glycoproteins
and is a gel-like substance.
• Different types of GAGs present are as follows: Chondroitin sulphate, hyaluronic acid, dermatan sulphate,
heparin, heparan sulphate and keratan sulphate.
• They are long chain un-branched polysaccharides.
• Proteoglycans are large molecules with a core protein in the centre and numerous GAGs (except hyaluronic
acid) attached around it.
o Hyaluronic acid is bound to core protein through a link protein.
• Glycoproteins have oligosaccharide chains attached to them.
o Types of glycoproteins: Fibronectin, laminin, entactin, osteonectin, chondronectin
o Fibronectin helps in adhesion of cells to extracellular matrix (ECM).
o Laminin helps in adhesion of epithelial cells to basement membrane (BM).
o Entactin binds laminin type IV collagen to BM.
o Osteonectin is present in bone and helps in bone mineralization
o Chondronectin is present in cartilages and helps in adhesion of chondrocytes to ECM.

FIBRES
• Fibres are formed by fibroblasts and are of 3 types: Collagen, elastic, reticular.
• Collagen fibres provide tensile strength. Collagen fibres are eosinophilic and appear pink on H & E stain. Some
types of collagen present in body are as follows:
o Type I: Presents in fascia, tendon, ligaments, etc.
o Type II: Presents in hyaline cartilage
o Type III: Presents in reticular fibres
o Type IV: Presents in basement membrane
• Elastic fibres can stretch and so provide elasticity. They are made of elastin protein and microfibrils.
o They are thinner than collagen fibres and show branching.
o They are eosinophilic and have the property to recoil.
• Reticular fibres form a network for framework in lymph nodes, spleen, bone marrow, basement membrane, etc.
o They are not stained by H & E stain.
Practical Manual of Histology 49

Three points of identification

Practical Manual of Histology 53

Date:

Hyaline cartilage
54 Practical Manual of Histology

Date:

ELASTIC CARTILAGE

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify elastic cartilage under light microscope.
2. Identify perichondrium and chondrocytes under light microscope.
3. Identify elastic fibres.
4. Draw a diagram of microanatomy of elastic cartilage.

Distribution and function

Three points of identification

Practical Manual of Histology 55

Date:

Elastic cartilage
56 Practical Manual of Histology

Date:

FIBROCARTILAGE

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify fibrocartilage under light microscope.
2. Identify chondrocytes under light microscope.
3. Give examples of fibrocartilage.
4. Draw a diagram of microanatomy of fibrocartilage.

Distribution and function

Three points of identification

Practical Manual of Histology 57

Date:

Fibrocartilage (intervertebral disc)

58 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 59

Date:

BONE

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 71.1 Identify bone under the K/S SH Y Lecture, practical Written/skill
microscope; classify various assessment
types and describe structure–
function correlation of the same

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify and describe the features of an LS of compact bone.
2. Identify and describe the features of a TS of compact bone.
3. Describe Haversian system.
4. Identify and describe the features of a TS of spongy bone.
5. Describe the histological components of bone.

HISTOLOGICAL COMPONENTS OF BONE

• Periosteum covers outer surface of bone. It has two layers—outer fibrous layer of collagen fibres; and inner
osteogenic layer having osteoprogenitor cells which form osteoblasts. Collagen fibres of outer layer binding
the periosteum to bone are called Sharpey’s fibres.
• Endosteum lines the inner surface of bone. It has a single layer of osteoprogenitor cells.
• Osteoprogenitor cells are irregular elongated cells having pale nucleus and a pale cytoplasm.
• Osteoblasts are large cells, cuboidal or low columnar, with cytoplasmic processes. They have intensely basophilic
cytoplasm and round nucleus situated at one side of cytoplasm and prominent nucleolus. They synthesize
collagen fibres and matrix of a bone.
• Osteocytes are flattened cells smaller than osteoblasts. They have processes that lie in canaliculi. Their cytoplasm
is less basophilic. They are present in the lacunae of bones. They are resting living cells. They can convert to
osteoblasts or osteoclasts.
• Osteoclasts are multinucleated giant cells with lightly basophilic or acidophilic cytoplasm. They are present on
bone surfaces that are undergoing resorption. They lie in Howship’s lacunae.
• Ground substance has organic and inorganic elements.

Practical Manual of Histology 65

Date:

TS of cancellous (spongy) bone

66 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 67

Date:

MUSCULAR TISSUE

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 67.1 Describe and identify various K/S SH Y Lecture, practical Written/skill
types of muscle under the assessment
microscope

SKELETAL MUSCLE (STRIATED, SOMATIC, STRIPED, VOLUNTARY)

• Muscle fibres are long, cylindrical and multinucleated. Nuclei are present at the periphery.
• Each fibre is covered by endomysium. Many muscle fibres are held together as fascicles. Each fasciculus is
surrounded by epimysium. Whole skeletal muscle is surrounded by perimysium.
• Muscle fibres have transverse and longitudinal striations. There are alternate light (I) and dark (A) bands in
transverse striations.
• There are neuromuscular spindles within the skeletal muscle.

SMOOTH MUSCLE (UNSTRIPED, INVOLUNTARY, UNSTRIATED)

Date:

TS of smooth muscle
78 Practical Manual of Histology

Date:

MUSCLE (ULTRASTRUCTURE)

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 67.2 Classify muscle and describe K/S SH Y Lecture, practical Written
the structure–function
correlation of the same
AN 67.3 Describe the ultrastructure K KH N Lecture, practical Written
of muscular tissue

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Classify muscle.
2. Correlate function with structure of muscle.
3. Describe ultrastructure of skeletal muscle.
4. Describe ultrastructure of smooth muscle.
5. Describe ultrastructure of cardiac muscle.

Answer the Following

Skeletal Muscle
1. Alternate light and dark bands in myofibrils are caused by a difference in the

2. Dark band is known as band (anisotropic).

3. Light band is known as band (isotropic).
4. A band possesses a light zone called line or Hensen’s line. In the middle of this light zone
there is a dark line called line.
5. I band has a dark line dividing it known as line or membrane.
6. There are 2 types of myofilaments in a myofibril known as and .
7. Sarcomere is a unit of contraction and lies between two lines.
8. Draw a schematic diagram of ultrastructure of a sarcomere.
Practical Manual of Histology 79

1. Draw the ultrastructure of body of neuron.

2. Draw a longitudinal section of ultrastructure of myelinated nerve fibres.

82 Practical Manual of Histology

Date:

PERIPHERAL NERVE (TS)

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify TS of a peripheral nerve under light microscope.
2. Identify the epineurium, perineurium and endoneurium.
3. Explain what myelin sheath looks like in H & E stain.
4. Draw a diagram of microstructure of TS of a peripheral nerve.

Distribution and function of Schwann cells

Three points of identification

Practical Manual of Histology 83

Date:

TS of peripheral nerve
84 Practical Manual of Histology

Date:

SENSORY/SPINAL/DORSAL ROOT GANGLION

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify a dorsal root ganglion under light microscope.
2. Identify the cell bodies of neurons and satellite cells.
3. Appreciate the clusters of cell bodies.
4. Identify whether it is unipolar or multipolar neuron.
5. Draw a diagram of microstructure of sensory ganglion.

Distribution and function

Three points of identification

Practical Manual of Histology 85

Date:

Dorsal root ganglion

86 Practical Manual of Histology

Date:

AUTONOMIC/SYMPATHETIC/PARASYMPATHETIC GANGLION

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify sympathetic ganglion under the light microscope.
2. Identify the multipolar neuron bodies.
3. Appreciate the wide spacing between cell bodies.
4. Appreciate the lesser number of satellite cells.
5. Draw the microstructure of an autonomic ganglion.

Distribution and function

Three points of identification

Practical Manual of Histology 87

Date:

Parasympathetic ganglion/autonomic ganglion

88 Practical Manual of Histology

Date:

EXOCRINE (SALIVARY) GLANDS HISTOLOGY

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 70.1 Identify exocrine gland under K/S SH Y Lecture, practical Written/skill
the microscope and distinguish assessment
between serous, mucous and
mixed acini

CLASSIFICATION BASED ON TYPE OF SECRETION

1. Mucous glands
2. Serous glands
3. Mixed glands

MICROSCOPIC STRUCTURE OF EXOCRINE GLANDS

• Parenchyma: It consists of secretory cells. Shape of secretory units of salivary glands is acinar (round or oval).
Therefore, there are serous, mucous or mixed acini depending upon the type of salivary gland.
• Stroma: It consists of connective tissue dividing the gland into lobules, lobes or forming its capsule. Blood
vessels pass through the connective tissue.
• Ducts: Intralobular, interlobular, interlobar
• Myoepithelial cells are present in close relation to the secretory elements. They are stellate or fusiform in shape
and lie between the secretory cells and basement membrane.

ACINI
• Mucous acini: They contain mucopolysaccharide secretion which does not stain with H & E stain. Therefore,
the cells of mucous acini appear empty. The secretions near the apices push the nuclei towards the base of cells
(near the basement membrane) making them appear flattened. There is a lumen in the centre of acinus which is
larger as compared to that of serous acinous, e.g. sublingual salivary gland.
• Serous acini: Cells are triangular with round nucleus placed centrally/basal. Secretions are proteinous, therefore,
the cytoplasm is granular and stains bluish with H & E. There are basal striations present in active cells in
higher magnification. Lumen is small, e.g. parotid gland.
• Mixed acini: Submandibular gland is a mixed salivary gland having both serous and mucous acini. Demilunes
are present on mucous acini.

Demilunes of Gianuzzi
The mucous cells form tubules, but their ends are capped by serous cells, which constitute the serous demilunes.
Under the light microscope the serous demilunes appear as semilunar dark stained caps on the light stained
mucous acini. The demilunes are connected with the lumen of mucous acini by microscopic canaliculi to the
lumen of acini. So the serous secretion of demilunes makes the secretion of mucous acini less viscid.

Ducts
• Intercalated ducts: Lined by cuboidal epithelium, are present in the lobules.
• Striated ducts: Formed by joining of several intercalated ducts. They are lined by simple columnar cells and
have basal striations formed by folding of basal cell membrane. They have a larger lumen than intercalated
ducts.
• Interlobular ducts: The epithelium varies from low columnar in smaller ducts to pseudostratified or stratified
columnar epithelium in larger ducts.
Practical Manual of Histology 89

Practical Manual of Histology 93

Date:

Parotid salivary gland (serous gland)

94 Practical Manual of Histology

Date:

SUBMANDIBULAR SALIVARY GLAND (MIXED GLAND)

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify mixed salivary gland under light microscope.
2. Identify the mucous and serous acini, ducts, connective tissue.
3. Identify the serous demilunes.
4. Draw the microanatomy of mixed salivary gland.

Distribution and function of mixed glands

Three points of identification

Practical Manual of Histology 95

Date:

Submandibular salivary gland (mixed gland)

96 Practical Manual of Histology

Date:

BLOOD VESSELS HISTOLOGY

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 69.1 Identify elastic and muscular K/S SH Y Lecture, practical Written/skill
blood vessels, capillaries under assessment
the microscope
AN 69.2 Describe the various types and K KH Y Lecture, practical Written
structure–function correlation
of blood vessels
AN 69.3 Describe the ultrastructure of K KH Y Lecture, practical Written
blood vessels

THINK AND ANSWER

1. Large arteries (like aorta, brachiocephalic trunk, common carotid, common iliac, etc.) receive lots of blood
under pressure in each systole. Then the blood passes forwards and the wall of arteries recoil. So what kind of
tissue should be present in the wall of large blood vessels?
2. You already know the epithelium lining of the lumen. What is it?
3. If a blood vessel can constrict or dilate then which tissue will be responsible for this?

Elastic Artery
• The diameter of the lumen is usually more than the thickness of the wall.
• It consists of 3 layers: Tunica intima, tunica media and tunica adventitia.
• Tunica intima has endothelium (of simple squamous epithelium), sub-endothelial connective tissue and internal
elastic lamina. The internal elastic lamina is not well developed in elastic artery.
• Tunica media is the thickest layer and mainly has elastic fibres apart from a few smooth muscle fibres.
• Tunica adventitia has collagenous connective tissue fibres with vasa vasorum and autonomic nerves.

Muscular Arteries
• The diameter of the lumen is usually less than the thickness of the wall. Layers are the same as elastic artery.
The difference is that the tunica intima shows folding due to the prominent internal elastic lamina which is
wavy, formed of condensed elastic fibres.
• The tunica media has concentrically arranged smooth muscle fibres. Very few elastic fibres are present.

Vein
• Wall of veins is thin and lumen is collapsed. Small and medium-sized veins have valves.
• The 3 layers are the same with the following differences:
o Internal elastic lamina is absent in tunica intima.
o Tunica media is thin and has very few elastic tissue and muscle fibres.
o Tunica adventitia is well developed and thicker than the tunica media. It has collagen fibres, smooth muscle
fibres, and a few elastic fibres.

Capillaries
• The wall of capillary is formed of a single layer of endothelial cells and basal lamina. Occasionally a pericyte
may be seen outside the basal lamina of continuous capillaries. Pericyte is a contractile cell.
Practical Manual of Histology 97

Date:

Ultrastructure of Capillaries
Capillaries are of 3 types:
1. Continuous capillary—here there are no fenestrations in the endothelial cells or basal lamina, e.g. capillaries of
muscles, nervous tissue, etc.
2. Fenestrated capillary—fenestrations of 70–90 micrometer diameter are present in the endothelium, through
which passage of fluid or blood components occurs. Basal lamina is continuous, e.g. capillaries of small intestine,
glomeruli, endocrine glands, etc.
3. Sinusoidal capillaries—these are large and irregular in shape. Endothelium is fenestrated and basal lamina is
also not continuous, e.g. capillaries of liver, spleen, bone marrow.

Draw the Microanatomy of the Given Capillary and Vein with Proper H & E Staining and Labelling

CAPILLARY VEIN
Draw a capillary according to the
description given above
98 Practical Manual of Histology

Date:

ELASTIC ARTERY/LARGE ARTERY

SPECIFIC LEARNING OBJECTIVES

Dense lymphoid tissue is an aggregation of lymphocytes which are arranged in the form of nodules. Therefore,
the microscopic appearance is similar to lymphocytes in H & E stain apart from the connective tissue within and
outside the tissue and its blood vessels. The scope of this book deals with structures mentioned in the competencies
above.

Think and Answer

1. What will be the colour taken up by the predominant cells (lymphocytes) in lymphoid tissue if stained by
H & E?

2. What will be the colour of connective tissue fibres within the tissue?

3. What is the function of spleen? So what do you expect to see in its microanatomy?

4. What is the function of thymus? So what kind of cells do you expect to see under the light microscope?

5. Does the thymus persist till adulthood? If not, then do you expect to see some degenerative changes under the
light microscope?

6. Where are palatine tonsils situated? So what kind of epithelium should be present on the side facing the oral
cavity?
Practical Manual of Histology 103

Date:

LYMPH NODE
• It has an outer zone of densely packed lymphocytes in the form of aggregates called lymphoid follicles or
lymphoid nodules. The nodule has a densely packed dark (basophilic) stained peripheral region and a paler
staining germinal centre.
• The central zone, medulla, is light stained with lymphocytes arranged as cords.
• Lymph node is surrounded by a capsule made up of mainly collagen fibres. The capsule sends septae (trabeculae)
within the node to divide into lobules.
• There is subcapsular sinus just below the capsule. It contains lymph which enters the LN through convex side
of LN. Then this lymph flows through the sinuses present along the trabeculae called cortical sinuses. This
reaches the medulla where there are medullary sinuses which join to form efferent lymphatic vessels which
emerge through the hilum.
• Blood vessels enter the hilum and pass through the medulla to reach the cortex.

Now Answer these Questions

1. What stain will lymph take on H & E staining?

2. So what will be the colour of sinuses?

3. What is the characteristic of lining epithelium of sinuses?

4. Where do you expect to see the blood vessels in lymph node?

104 Practical Manual of Histology

Date:

LYMPH NODE

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify lymph node under light microscope.
2. Identify the lymphatic follicles, cortex and medulla.
3. Identify the subcapsular and medullary sinuses.
4. Identify the capsule and trabeculae.
5. Draw the microanatomy of lymph node.

Distribution and function

Three points of identification

Practical Manual of Histology 105

Date:

Lymph node
106 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 107

Date:

SPLEEN
• Outermost is a layer of mesothelium (Can you tell what kind of epithelium is present in mesothelium?
). Spleen is surrounded by dense connective tissue-capsule. It sends trabeculae inside the
substance of spleen. Blood vessels enter along the trabeculae.
• The arrangement of lymphoid tissue in spleen is not like cortex and medulla. Instead it is arranged as red pulp
and white pulp.
• The characteristic reticular fibre network of spleen is not visible in H & E staining.

WHITE PULP
• The arteries enter the parenchyma as arterioles and are surrounded by a sheath of T lymphocytes. This sheath
is known as periarterial lymphatic sheath (PALS). So these will appear as cords because the arteriole extends in
length and it is surrounded by PALS. If cut in TS, it appears round with central artery.
• At places the arrangement of lymphocytes becomes nodular and these are known as lymphatic nodules. These
nodules have B lymphocytes with a light stained and loosely packed germinal centre surrounded by densely
packed dark staining peripheral lymphocytes. These nodules are known as malpighian bodies.
• There is a central arteriole which is eccentric in position. This arteriole usually lies between the germinal centre
and densely packed lymphocytes. This is the characteristic and differentiating feature of lymphatic nodule of
spleen.

RED PULP
• This is another characteristic feature of microscopic structure of spleen. The red pulp consists of all cells of
blood. There are sinusoids (can you tell the lining epithelium of sinusoids? ) which are
surrounded by cords of blood cells (T lymphocytes, B lymphocytes, macrophages, other cells of blood). These
cords are known as cords of Billroth. The red pulp surrounding the lymphatic nodule is made of sinusoids and
is known as marginal zone.

SPLENIC CIRCULATION
108 Practical Manual of Histology

Date:

SPLEEN

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify spleen under light microscope.
2. Identify red pulp and white pulp.
3. Identify central artery in white pulp.
4. Identify medullary cords and sinuses.
5. Draw a diagram of microanatomy of spleen.

Function of red and white pulps

Three points of identification

Practical Manual of Histology 109

Date:

Spleen
110 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 111

Date:

THYMUS
• Thymus is divided into incomplete lobules by septae from capsule.
• Thus there is cortex in the periphery and medulla in the centre of each lobule.
• Medulla is placed in the centre and communicates with the medulla of adjoining incomplete lobules.
• Cortex of thymus contains densely packed T lymphocytes, epithelioreticular cells (epitheliocytes) and
macrophages.
• Medulla contains fewer lymphocytes but more epitheliocytes. Concentrically arranged degenerating
epitheliocytes form Hassall’s corpuscles or thymic corpuscles (characteristic feature of thymus).
• Composition of Hassall’s corpuscles: Degenerating epitheliocytes, lipid and macrophages.

Answer the Following

1. What are Hassall’s corpuscles?

2. What are the changes in thymus gland with advancing age?

112 Practical Manual of Histology

Date:

THYMUS

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify thymus under the light microscope.
2. Identify its lobules and cortex and medulla.
3. Identify the lymphoid cells.
4. Identify Hassall’s corpuscles.
5. Draw a diagram of microanatomy of thymus.

Distribution and Function of thymus

Three points of identification

Practical Manual of Histology 113

Date:

Thymus gland
114 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 115

Date:

PALATINE TONSIL
• Tonsils are collections of lymphoid tissues.
• Palatine tonsil is present in the junction of oral cavity with pharynx. As such the covering epithelium is same
as that of the oral cavity, i.e. stratified squamous non-keratinized.
• Tonsillar crypts are seen as extensions of covering epithelium within the substance of the tonsil.

Revise and Answer

1. What are the features of stratified squamous non-keratinized epithelium?

2. How can you correlate tonsillar crypts with development of palatine tonsils?

3. What kind of muscles fibres (smooth/skeletal) may be seen on the deeper side of palatine tonsils?

4. Name the other tonsils participating in the formation of Waldeyer’s ring.

116 Practical Manual of Histology

Date:

PALATINE TONSIL

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify palatine tonsil under the light microscope.
2. Identify lymphoid nodules in the tissue.
3. Identify stratified squamous epithelial lining of the tissue.
4. Identify tonsillar crypts.
5. Identify muscle fibres in the deeper side of palatine tonsil (if present in section).
6. Draw a diagram of microanatomy of palatine tonsil.

Function of lymphoid nodules

Three points of identification

Practical Manual of Histology 117

Date:

Palatine tonsil
118 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 119

Date:

INTEGUMENTARY SYSTEM

(Y/N) method method to certify
AN 25.1 Identify, draw and label a K/S SH Y Lecture, practical Written/skill 1
slide of trachea and lung assessment

TRACHEA
• It has pseudostratified ciliated columnar epithelium with goblet cells resting on a basement membrane.
• Lamina propria has elastic fibres, WBCs and ducts of glands.
• Submucosa has both mucous and serous acini.
• C-shaped hyaline cartilage lies beneath the submucosa. The posterior gap in the cartilage has smooth muscle
fibres known as trachealis.
• Adventitia is the outermost layer.

Answer the Following

1. What type of cartilage is present in the trachea? What is its shape?

2. Why does the lumen of trachea not collapse?

3. How are chondrocytes arranged in the cartilage—groups or single?

4. What is the outer lining of cartilage known as?

5. What is the appearance of ground substance of hyaline cartilage in H & E stain and why?

6. What kind of epithelium lining do you expect for trapping foreign particles in the inspired air?

7. What is the lining epithelium of trachea?

8. What is the nature of secretion by glands in trachea?

126 Practical Manual of Histology

Date:

TRACHEA

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify trachea under the light microscope.
2. Identify the pseudostratified ciliated columnar epithelium with goblet cells.
3. Identify the hyaline cartilage, perichondrium, chondrocytes, lacunae and homogenous ground substance.
4. Identify the mucous and serous glands between cartilage and epithelium.
5. Identify smooth muscle—trachealis
6. Draw a diagram of microanatomy of trachea.

Distribution and function of pseudostratified epithelium

Three points of identification

Practical Manual of Histology 127

At the end of the session the student should be able to:
1. Identify epiglottis under the light microscope.
2. Identify stratified squamous non-keratinized epithelium.
3. Identify elastic cartilage.
4. Identify lamina propria and seromucinous glands within it.
5. Identify pseudostratified columnar epithelium.
6. Draw a diagram of microanatomy of epiglottis.

Distribution and function of elastic cartilage

Three points of identification

Practical Manual of Histology 137

Date:

Epiglottis (TS)
138 Practical Manual of Histology

Date:

GASTROINTESTINAL SYSTEM

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 52.1 Describe and identify the K/S SH Y Lecture, practical Written/skill
microanatomical features of assessment
Gastrointestinal system:
Oesophagus, fundus of stomach,
pylorus of stomach, duodenum,
jejunum, ileum, large intestine,
appendix, liver, gall bladder,
pancreas and suprarenals gland

General Layout of GIT

• There are four layers: Mucosa, submucosa, muscularis externa, serosa or adventitia.
• Mucosa is the lining epithelium along with lamina propria and muscularis interna. Lamina propria is connective
tissue with blood supply, lymphatics, glands and lymphoid follicles.
• Submucosa consists of loose connective tissue. It may have glands also.
• Muscularis externa has two layers of smooth muscles—inner circular and outer longitudinal.
• Adventitia is loose connective tissue which is covered with mesothelium in the abdomen.
• Adventitia + mesothelium = Serosa.
Practical Manual of Histology 139

Date:

TONGUE

Number Competency Domain Level Core Teaching/learning Assessment Number required

Think and Answer

1. Can you move the tongue at your will? So what kind of muscle will be found in the tongue?

2. What is the special sense perceived by the tongue? So what will be found in histology?

3. What is the epithelial lining of oral cavity? Why?

• The oral cavity is lined by stratified squamous epithelium which is non-keratinized in most places. In the
tongue the filiform papillae show keratinisation due to frequent mechanical stress.
• There is lamina propria but submucosa is absent in tongue. Lamina propria has minor salivary glands which
are serous and mucous glands.
• The skeletal muscles of tongue run in different directions. So on section the fibres can be seen in different
directions and sections as bundles.
• Taste buds are seen on circumvallate and fungiform papillae of tongue. Taste buds have oval cells arranged in
clusters. At the apex of the taste bud there is a taste pore. The portion of cells towards the taste pore has
microvilli.
140 Practical Manual of Histology

Date:

TONGUE

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify tongue under the light microscope.
2. Identify its epithelium (stratified squamous non-keratinized).
3. Identify its papillae and taste buds.
4. Identify its skeletal muscles running in different directions.
5. Identify minor salivary glands.
6. Draw a diagram of microanatomy of tongue.

Distribution and function of taste buds and filiform papillae

Three points of identification

Practical Manual of Histology 141

Date:

Tongue (circumvallate papillae)

142 Practical Manual of Histology

Date:

Tongue (fungiform and filiform papillae)

Practical Manual of Histology 143

Date:

OESOPHAGUS
The four layers are the same as the general plan of GIT.
1. Mucosa has stratified squamous non-keratinized epithelium. Lamina propria may have lymphoid follicles.
Muscularis interna has smooth muscles.
2. Submucosa has mucous glands, blood vessels, lymphatics and Meissener’s plexus of nerves.
3. Muscularis externa in the upper 1/3rd of oesophagus there are skeletal muscles, in middle 1/3rd there are
smooth and skeletal muscles and lower 1/3rd there are only smooth muscles.
4. Outermost layer consists of adventitia in the upper part of oesophagus and serosa in abdominal part.

Three points of identification

Practical Manual of Histology 149

Date:

TS through middle one-third of oesophagus

TS of oesophagus through middle one-third

Hint: In muscularis externa draw TS of both smooth and skeletal muscles

150 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 151

Date:

CARDIOESOPHAGEAL JUNCTION

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 52.3 Describe and identify the K/S SH Y Lecture, practical Written/skill
microanatomical features of assessment
cardioesophageal junction,
corpus luteum

• Cardioesophageal junction is the junction of the upper end of stomach (cardiac end) and lower end of
oesophagus.
• The stratified squamous epithelium of oesophagus abruptly changes into simple columnar epithelium. There
are no goblet cells.
• Gastric pits lined by simple columnar cells are present.
• Lamina propria has tubular glands having columnar cells with a few parietal and zymogenic cells at the base
of gland.
• Muscularis mucosa has inner circular and outer longitudinal smooth muscle layers.
• Submucosa may have mucous acini apart from loose connective tissue.
• Muscularis externa has inner circular and outer longitudinal smooth muscle layers.
• It is covered by serosa (simple squamous epithelium).
152 Practical Manual of Histology

Date:

CARDIOESOPHAGEAL JUNCTION

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify cardioesophageal junction under the light microscope.
2. Identify the junction of stratified squamous non-keratinized epithelium and simple columnar epithelium.
3. Identify the four layers: Mucosa, submucosa, muscularis externa and serosa.
4. Identify gastric pits and their cells in lamina propria.
5. Identify mucous glands in submucosa.
6. Draw a diagram of microanatomy of cardioesophageal junction.

Applied importance of cardioesophageal junction

Three points of identification

Practical Manual of Histology 153

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Cardioesophageal junction
154 Practical Manual of Histology

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NOTES
Practical Manual of Histology 155

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STOMACH

TS THROUGH FUNDUS/BODY OF STOMACH

Fundus and Body of Stomach
• Histologically, the fundus and body of stomach are similar.
• Apart from the general layout of GIT, the surface epithelium dips into the lamina propria to form gastric pits.
A few (3–7) gastric glands open into each gastric pit (lined by columnar cells).
• Base and body of gastric gland is lined by chief cells/zymogenic/peptic cells. They are pyramidal in shape and
have a round nucleus in the centre with granular cytoplasm which stains blue (basophilic). They secrete enzymes.
• Between the chief cells are occasional oxyntic/parietal cells. They secrete H+ and Cl– and intrinsic factor. They
characteristically appear as large pink cells with large oval nuclei. So they are bright acidophilic cells. These
large cells bulge in the lumen of gastric pit and give a beaded appearance.
• Mucous neck cells are present as lining the neck of gastric gland. They are low columnar cells, have mucous
and round nuclei.
• Argentaffin cells (triangular cells) at the base of the gastric gland are not stained by H & E staining and require
silver salts for staining.
• Muscularis mucosa has thin inner circular and outer longitudinal smooth muscle layers.
• The muscularis externa of body and fundus is different from the general plan of two layers of GIT. It has three
layers—innermost oblique, middle circular and outer longitudinal.
• Serosa has squamous cells.

Think and Answer

a. What kind of epithelium will you make in microanatomy of fundus of stomach?

b. What all will you draw in lamina propria of fundus of stomach?

c. How will you draw the muscularis mucosae?

d. What will you draw in submucosa?

e. How many layers of muscles will you draw in muscularis externa of fundus/ body of stomach? How will you
draw the different layers?

f. What will be the outermost covering of fundus of stomach?

156 Practical Manual of Histology

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FUNDUS/ BODY OF STOMACH (TS)

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify the slide under light microscope.
2. Identify the type of epithelium lining the fundus of stomach.
3. Identify the chief cells and oxyntic cells (beaded appearance).
4. Identify the gastric pits and glands.
5. Identify the muscularis mucosa, submucosa, muscularis externa and serosa.
6. Draw a diagram of microanatomy of fundus of stomach.

Function of chief and parietal cells

Three points of identification

Practical Manual of Histology 157

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Fundus/body of stomach (TS)

158 Practical Manual of Histology

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NOTES
Practical Manual of Histology 159

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TS THROUGH PYLORUS OF STOMACH

Points of differences between histology of pylorus and fundus/body of stomach
1. Gastric pits are deeper and pyloric glands are shorter with larger lumen.
2. Acini of glands are lightly stained columnar cells, but the nuclei are flattened and situated in the periphery.
They secrete mucous and hormone gastrin. They are situated in the lamina propria (same as fundus)
3. Some oxyntic cells may be present.
4. Muscularis externa consists chiefly of thickened circular layer forming the pyloric sphincter.

Now Answer the Following

a. What will be the lining epithelium of pylorus of stomach?

b. What is the characteristic of acini of pyloric glands?

c. What layers of stomach wall will you make?

d. How many layers of muscularis externa will be drawn? Name them.

160 Practical Manual of Histology

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TS THROUGH PYLORUS OF STOMACH

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify pylorus under the light microscope.
2. Identify all the layers of GIT in the wall of pylorus.
3. Identify pyloric glands
4. Identify thick circular layer of smooth muscles of muscularis externa.
5. Draw a diagram of microanatomy of pylorus.

Function of inner circular layer of muscularis externa in pylorus of stomach

Three points of identification

Practical Manual of Histology 161

Date:

TS through pylorus of stomach

162 Practical Manual of Histology

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NOTES
Practical Manual of Histology 163

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INTESTINES

SMALL INTESTINES
• Mucosa—it has permanent folds known as valves of Kerckring. They can be seen with the naked eye. They
have mucosa and submucosa.
• Mucosa has intestinal villi which have a core of lamina propria surrounded by columnar epithelium.
• Intestinal glands known as crypts of Lieberkuhn which lie in the lamina propria. They are invaginations of the
epithelium (columnar epithelium). They open at the bases of villi. Paneth cells may be seen in the deeper parts
of crypts as pink cells because of eosinophilic granules of lysozyme and antibacterial substance. They are
pyramidal in shape with basal nuclei
• Lymphocytes may be present as follicles.

DUODENUM
Distinctive Features of Duodenum
• Villi are broad and leaf-shaped with brush border and goblet cells.
• Inner layer of muscularis mucosae sends extensions into the villi. So smooth muscles will be visible in the villi.
• Submucosa has mucous glands known as Brunner’s glands or duodenal glands. These have columnar cells
with a flat nucleus at the base. Their ducts open into the crypts of Lieberkuhn. They secrete mucous which is
alkaline.

Now Answer the Following

LARGE INTESTINE

MICROSCOPIC FEATURES OF LARGE INTESTINE

• There are no villi or plica circulares as compared from the small intestine.
• The lining epithelium is simple columnar but has numerous goblet cells.
• Microvilli are present which make the outline of columnar cells appear dark pink stained.
• Deep crypts of Lieberkuhn are present. They also have goblet cells.
• Lamina propria has lymphoid follicles.
• Submucosa is the same as the general plan of GIT.
• Muscularis externa has the same two layers but the outer longitudinal layer forms three bands known as
taenia coli in addition to forming the outer layer.
• Serosa (mesothelium) + adventitia covering is seen in transverse and sigmoid colon, whereas retroperitoneal
ascending and descending colon have only adventitia covering.

Answer the Following

1. List the differences between small and large intestines.

Small intestine Large intestine

2. Which muscular coat forms taenia coli?

Practical Manual of Histology 171

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VERMIFORM APPENDIX

DIFFERENCES FROM LARGE INTESTINE

• Smaller lumen
• There are no taenia coli
• There are numerous lymphoid follicles in lamina propria which encroach into the submucosa.
• Muscularis mucosa is interrupted and muscularis externa is thin.
• Serosa covers it completely.

Answer the Following

1. How will you differentiate between appendix and fallopian tube under the light microscope?

2. What is the difference between lymphoid follicles of appendix and ileum under microscope?

3. What is the difference between lumen of appendix and vas deferens as seen in microscope?

4. How will you differentiate between the microstructure of appendix and lymph node?
172 Practical Manual of Histology

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TS THROUGH VERMIFORM APPENDIX

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify vermiform appendix under the light microscope
2. Identify lymphoid follicles in lamina propria and submucosa.
3. Identify the interrupted muscularis mucosae and thin muscularis externa.
4. Identify serosa covering.
5. Differentiate it from microscopic appearance of lymph node.
6. Draw a diagram of microanatomy of vermiform appendix.

Function of appendix

Three points of identification

Practical Manual of Histology 173

LIVER
• Liver is covered by Glisson’s capsule seen as eosinophilic connective tissue on H & E staining. This extends in
the substance of liver and divides it into the characteristic lobules.
• Classical lobule is hexagonal in shape. The hepatocytes in parenchyma are polyhedral in shape. They are
arranged in plates seen as radiating from the central vein. They are anastomosing and separated by hepatic
sinusoids. Space between hepatocytes and sinusoids is known as space of Disse which lodges hepatic stellate
cells called Ito cells (identified by light staining vacuolar appearance due to lipid droplets).
• Sinusoids are lined by phagocytic Kupffer cells (stellate/rounded in shape, difficult to identify under light
microscope) and endothelial cells (simple squamous epithelium).
• The angles of hexagon have portal triads consisting of branches of portal artery (lined by endothelium having
tunica media with smooth muscles and round lumen), portal vein tributaries (lined by endothelium and having
thin tunica media but lumen larger than the artery) and bile ductules (lined by simple cuboidal epithelium).
The centre has a central vein.
178 Practical Manual of Histology

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LIVER

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify liver under the light microscope.
2. Identify its capsule, hepatocytes and hepatic lobules.
3. Identify the portal triad and its component vessels—hepatic arteriole, hepatic venule, bile ductule.
4. Identify central vein, hepatic sinusoids.
5. Draw a diagram of microanatomy of liver.

Function of sinusoids of liver

Three points of identification

Practical Manual of Histology 179

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Liver
180 Practical Manual of Histology

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NOTES
Practical Manual of Histology 181

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PANCREAS
• Pancreas is covered by a thin layer of connective tissue. Numerous septa arise from this and divide the
parenchyma into lobules.
• Exocrine part has acini which are serous (like acini of parotid gland).
• Intercalated duct, intralobular duct (simple cuboidal), and interlobular ducts (simple columnar) are seen which
finally drain into pancreatic duct.
• Endocrine part has islets of Langerhans which are scattered between exocrine acini. Cells of islets are polyhedral
in shape and arranged as cords. Capillaries are seen between the polyhedral cells. Alpha, delta, gamma and PP
(pancreatic polypeptide cells which were formerly known as gamma cells) types of cells of islets of Langerhans
cannot be identified in H & E staining. This requires immune-cytochemical methods to be identified.
• The cells of islets stain light and appear pale. Alpha cells have red granules and beta cells have blue granules.
They are present in clusters.

Answer the Following

1. Which salivary gland has serous acini?

2. What is the shape of cell and colour of cytoplasm of serous acini?

3. Where is the nucleus located in a cell of serous acinus?

4. How will you differentiate a pancreas from parotid gland on light microscopy?
182 Practical Manual of Histology

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PANCREAS

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify pancreas under the light microscope.
2. Identify lobules and serous acini.
3. Identify islets of Langerhans and capillaries in it.
4. Draw a diagram of microanatomy of pancreas.

Function of serous acinic and islets of Langerhans in pancreas

Three points of identification

Practical Manual of Histology 183

Date:

Pancreas
184 Practical Manual of Histology

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NOTES
Practical Manual of Histology 185

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GALL BLADDER
• Gall bladder mucosa has temporary folds.
• Its epithelium is of tall columnar cells with nuclei oval in shape and near the base.
• There are no goblet cells in the epithelium.
• Luminal surface of the cells have a dark pink border as seen under the microscope. This is due to microvilli
which form brush border.
• Lamina propria is present.
• Muscularis mucosae and glands are absent.
• So outer to the lamina propria there is a fibromuscular layer which represents muscularis externa. So it will
have pink staining collagen fibres and spindle-shaped smooth muscles with pink cytoplasm and blue central
nucleus.
• Outermost layer is of serosa or adventitia.

Think and Answer

1. How will you differentiate the mucosal folds of gall bladder from the villi of small intestine?

2. What is the difference between muscularis externa of general GIT plan and that of gall bladder?

3. What is brush border?

186 Practical Manual of Histology

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GALL BLADDER

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify gall bladder under the light microscope.
2. Identify the lining epithelium of gall bladder.
3. Identify the absence of muscularis mucosae.
4. Identify the layers of wall of gall bladder.
5. Draw a diagram of microanatomy of gall bladder.

Function of gall bladder

Three points of identification

Practical Manual of Histology 187

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Gall bladder
188 Practical Manual of Histology

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URINARY SYSTEM

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN52.2 Describe and identify the K/S SH Y Lecture, practical Written/skill
microanatomical features of: assessment
Urinary system: Kidney, ureter
and urinary bladder
Male reproductive system:
Testis, epididymis, vas deferens,
prostate and penis
Female reproductive system:
Ovary, uterus, uterine tube, cervix,
placenta and umbilical cord

The various structures encountered in the kidney have been mentioned in the schematic diagrams given below.
It is advisable to go through each of the given structures and their microscopic characteristic mentioned along
with it.

Schematic diagram of LS of kidney

Practical Manual of Histology 189

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Schematic diagram of a nephron and collecting system showing microscopic characteristics on H & E staining

JUXTAGLOMERULAR APPARATUS
It is situated near the glomerulus. It has three components:
1. Macula densa is modified cells in the DCT near the afferent arteriole. The cells are columnar (instead of cuboidal
of DCT).
2. Juxtaglomerular cells are modified smooth muscle cells of the tunica media of afferent arterioles of glomerulus.
Their cells are large and rounded with spherical nuclei.
3. Lacis cells/cells of polkissen/extraglomerular mesangial cells. They are polyhedral cells between macula densa,
afferent and efferent arterioles and vascular pole of kidney.
190 Practical Manual of Histology

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KIDNEY

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify kidney under the light microscope.
2. Identify cortex having renal corpuscles, PCT (more numerous in number), DCT, and collecting ducts.
3. Indentify medulla having more of collecting ducts, loop of Henle and vasa recta.
4. Identify Bowman’s capsule, glomerulus.
5. Appreciate the brush border of PCT.
6. Draw a diagram of microanatomy of kidney.

Function of kidney

Three points of identification

Practical Manual of Histology 191

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Kidney
192 Practical Manual of Histology

1. How many layers of cells are there in epithelium of stretched bladder?

2. What happens to the cells when urinary bladder stretches?

3. Which layer of muscles will form the sphincter vesicae?

4. Why is the superior surface of bladder covered with serosa instead of adventitia?
198 Practical Manual of Histology

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URINARY BLADDER

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify urinary bladder under the light microscope.
2. Identify transitional epithelium.
3. Identify muscular layers.
4. Identify serosa/adventitia.
5. Draw the diagram of microanatomy of urinary bladder.

Function of transitional epithelium

Three points of identification

Practical Manual of Histology 199

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EPIDIDYMIS
• A TS through epididymis shows numerous tubules of different sizes.
• The lining of these tubules is of pseudostratified columnar epithelium.
• They rest on a basement membrane and have smooth muscle fibres arranged around the tubules.
• Luminal surface of the columnar cells has non-motile stereocilia.
• Lumen shows spermatozoa within it. (Function of epididymis is storage of sperms and helping in their
maturation.)
• Columnar cells absorb testicular fluid and remove degenerating sperms.

Revise Previous Chapters and Answer the Following

1. What is the characteristic feature of pseudostratified columnar epithelium?

2. How will you differentiate between pseudostratified columnar epithelium of respiratory system from that of
epididymis?

3. What is the role of smooth muscle fibres around the tubules?

4. What is the difference between stereocilia and cilia?

206 Practical Manual of Histology

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EPIDIDYMIS

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify epididymis under the light microscope.
2. Identify pseudostratified columnar epithelium of epididymis.
3. Identify stereocilia.
4. Identify spermatids in the lumen.
5. Draw a diagram of microanatomy of epididymis.

Function of stereocilia in epididymis

Three points of identification

Practical Manual of Histology 207

Date:

Epididymis
208 Practical Manual of Histology

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NOTES
Practical Manual of Histology 209

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VAS DEFERENS/DUCTUS DEFERENS

• It has a star-shaped lumen as seen under the light microscope.
• It has three layers: Inner mucous membrane, middle muscular coat, outer fibrous coat.
• Epithelium is pseudostratified columnar epithelium in the distal part and simple ciliated columnar in the
proximal part.
• Lamina propria has elastic fibres which cause the epithelium to be thrown into folds. So the lumen appears
star-shaped.
• Middle muscular coat is thick and has three layers: Inner and outer are longitudinal muscle fibres and middle
layer has circular muscle fibres. The inner longitudinal layer is present in the proximal part of vas deferens.
• Outermost is the adventitia with blood vessels and nerves.
• The whole vas deferens can be seen in one field in low power.

Think and Answer

1. What is the appearance of the shape of lumen of ureter under the light microscope?

2. How will you differentiate between a slide of ureter and vas deferens under the light microscope?

3. What are the examples of tissues with pseudostratified columnar epithelium?

210 Practical Manual of Histology

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VAS DEFERENS

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify vas deferens under the light microscope.
2. Identify the lining epithelium and star-shaped lumen.
3. Identify layers of muscular coat.
4. Draw a diagram of microanatomy of vas deferens.

Function of vas deferens

Three points of identification

Practical Manual of Histology 211

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Vas deferens (TS)

212 Practical Manual of Histology

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NOTES
Practical Manual of Histology 213

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PROSTATE
• Prostate is traversed by prostatic urethra. So this part of urethra has three layers: Mucosa, submucosa and
muscle layer. Mucosa has pseudostratified columnar epithelium. It has recesses in which mucosal glands open.
Submucosa has loose connective tissue and submucosal glands which have short ducts which open in the
urethra. The muscle coat consists of inner longitudinal and outer circular layer of smooth muscles. All this is in
the central zone of prostate.
• An adult prostate consists of tubuloalveolar glands in a fibromuscular tissue. These are main prostatic glands
found in the peripheral zone. They have long ducts lined by bilaminar epithelium having columnar cell layer
and cuboidal cell layer. These ducts open into the prostatic urethra.
• The glandular part is not well-developed before puberty and degenerates in old age.
• Glandular tissue appears as follicles lined by columnar epithelium. The epithelium is pseudostratified in active
glands. This epithelium is characteristically thrown into numerous folds. Near the base there are basal cells
responsible for regeneration of epithelium.
• Amyloid bodies/corpora amylacea are seen within the alveoli of follicles in older age. They appear as
eosinophilic, lamellated round masses. They are made of glycoprotein.
• The fibromuscular tissue has collagen fibres and smooth muscle. They form septa between glandular follicles.
• Prostate is surrounded by fibrous capsule having numerous veins and parasympathetic ganglion cells.
214 Practical Manual of Histology

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PROSTATE

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify prostate under the light microscope.
2. Identify glandular follicles lined by columnar epithelium.
3. Appreciate the numerous mucosal folds of glandular follicles.
4. Identify collagen fibres and smooth muscle fibres between follicles.
5. Identify corpora amylacea in the follicles.
6. Identify prostatic urethra, if present in section.
7. Draw a diagram of microanatomy of prostate.

Function of glandular follicles of prostate

Three points of identification

Practical Manual of Histology 215

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Prostate
216 Practical Manual of Histology

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NOTES
Practical Manual of Histology 217

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PENIS
• The outermost covering of penis is thin skin. Hair follicles are present at the root of penis, the rest of the penis
being devoid of hair follicles.
• The dermis has smooth muscles and dilated venous spaces.
• There are three cavernous tissues—two corpora cavernosa on the dorsal side and one corpus spongiosum on
the ventral side.
• Corpus cavernosum is surrounded by fibrous (collagen and elastic) covering known as tunica albuginea. There
are spaces (caverns) which are enclosed by connective tissues and smooth muscle fibres forming trabeculae.
Caverns are vascular spaces lined by endothelial cells. There are arterioles and venules.
• Corpus spongiosum has finer trabeculae and narrower arterioles. Throughout the length of corpus spongiosum
is the spongy urethra which traverses the penis. Spongy urethra is lined by pseudostratified columnar epithelium
except near the terminal part where it is lined by stratified squamous epithelium. The fibrous sheath surrounding
the corpus spongiosum is thinner.
• A common fibrous sheath surrounds the corpora cavernosa and corpus spongiosum.
218 Practical Manual of Histology

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PENIS

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify penis under the light microscope.
2. Identify corpora cavernosa and corpus spongiosum.
3. Identify the urethra in corpus spongiosum.
4. Identify tunica albuginea.
5. Identify the epidermis and dermis layers of skin of penis.
6. Identify the smooth muscle fibres in dermis.
7. Draw the diagram of microanatomy of penis.

Function of corpora cavernosa and corpus spongiosum

Three points of identification

Practical Manual of Histology 219

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Penis (TS)
220 Practical Manual of Histology

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NOTES
Practical Manual of Histology 221

Date:

FEMALE REPRODUCTIVE SYSTEM

OVARY
• Ovary is the organ responsible for production of ova and hormones of female reproductive system.
• The germinal epithelium producing primordial follicles and ultimately ova is derived from its epithelial covering.
This means that the serosal covering of ovary is modified to cuboidal epithelium to form the germinal epithelium.
• The outer cortex of ovary has follicles in different stages of development, viz. primordial follicle, primary
follicle, secondary follicle, Graafian follicle, corpus luteum and atretic follicles.
• Apart from the follicles cortex has highly cellular connective tissue stroma.
• The inner medulla has loose connective tissue containing blood vessels, elastic fibres and a few smooth muscle
fibres.
Histological appearance of the various stages is as follows:
• Primordial follicle:
o Primary oocyte surrounded by simple squamous epithelium.
• Primary follicle:
o Larger oocyte surrounded by cuboidal epithelium. Between the two is the zona pellucida.
o On further development the cuboidal follicular cell layer becomes multilayered. These follicular cells are
called granulosa cells. They are surrounded by a thick basal lamina which appears eosinophilic on H & E
staining.
o Outside this lamina there are stromal cells of the ovary which differentiate into cuboidal cells layer—theca
interna and outer fibrous layer—theca externa.
• Secondary follicle:
o Initially numerous fluid filled cavities appear between the follicular cells. These fuse together to form a
single cavity called antrum. The liquid within it (liquor folliculi) is not stained by H & E staining, so it appears
empty.
o Granulosa cells at the side of antrum away from oocyte are known as membrana granulosa.
o Granulosa cells at the side of antrum adjacent to oocyte are known as cumulus oophoricus/cumulus ovaricus.
• Graafian follicle:
o The features of mature Graafian follicle as seen under the microscope are the same as secondary follicle with
single antrum with the following differences:
– Antrum is much larger.
– The feature not visible under the light microscope is the completion of 1st meiotic division and formation
of secondary oocyte. So the oocyte is now known as secondary oocyte.
222 Practical Manual of Histology

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OVARY

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify ovary under the light microscope.
2. Identify germinal epithelium.
3. Identify cortex and medulla.
4. Identify primordial, primary and secondary follicles.
5. Identify Graafian follicle and its components.
6. Identify corpus luteum, if present.
7. Draw a diagram of microanatomy of ovary.

Function of Graafian follicle

Three points of identification

Practical Manual of Histology 223

Date:

Ovary
224 Practical Manual of Histology

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NOTES
Practical Manual of Histology 225

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UTERUS
• The wall of uterus has three layers from within outwards: Endometrium, myometrium and serosa or adventitia.
• Endometrium has simple columnar epithelium with lamina propria underneath it.
• Lamina propria has simple tubular glands called endometrial glands.
• The deeper basal parts of these glands are not shed in menstruation. This part is called stratum basale. The
superficial part of endometrium is called stratum functionale which is shed during menstruation.
• Myometrium is the thickest layer of uterine wall. The bundles of smooth muscle fibres run in different directions.
There are blood vessels present between the bundles of smooth muscle.
• Adventitia (on the part of uterus not covered by peritoneum) or serosa (on the part of uterus covered by
peritoneum) is the outermost layer.
Proliferative phase (from 4th to 14th day of menstrual cycle)
• Stratum functionale regenerates from stratum basale.
• Thickness of stratum functionale increases.
• Length of endometrial glands increases and spiral artery also increases in length.
Secretory phase (14th to 28th day of menstrual cycle)
• Thickness of endometrium becomes double of proliferative phase.
• Glands become coiled giving the ‘saw tooth appearance’ in sections. The spiral arteries become coiled and
increase in length and span the whole length of stratum functionale.
Menstrual phase (1st to 4th day of menstrual cycle)
• The whole stratum functionale is shed.
• Stratum basale and its arteries remain.
226 Practical Manual of Histology

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UTERUS (PROLIFERATIVE PHASE)

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify uterus (proliferative phase) under the light microscope.
2. Identify epithelium and its columnar epithelium.
3. Identify lamina propria and tubular endometrial glands.
4. Identify myometrium and blood vessels in it.
5. Draw a diagram of microanatomy of proliferative phase of uterus.

Function of deciduas basalis

Three points of identification

Practical Manual of Histology 227

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Uterus (proliferative phase)

228 Practical Manual of Histology

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UTERUS (SECRETORY PHASE)

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify uterus (secretory phase) under the light microscope.
2. Identify endometrium, myometrium, adventitia/serosa.
3. Identify coiled tubular endometrial glands by their ‘saw-tooth appearance’.
4. Appreciate the greater thickness of endometrium.
5. Identify arteries between muscle bundles and lamina propria.
6. Draw a diagram of microanatomy of secretory phase of uterus.

Function of endometrial glands

Three points of identification

Practical Manual of Histology 229

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Uterus (secretory phase)

230 Practical Manual of Histology

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NOTES
Practical Manual of Histology 231

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CERVIX
• A section of cervix shows mucosal epithelium, lamina propria and connective tissue.
• Mucosal epithelium of cervical canal is simple columnar.
• There are cervical glands in lamina propria which open on the epithelium.
• Beyond the external os the simple columnar epithelium changes to non-keratinized squamous epithelium and
continues over the outer surface of vaginal part of cervix.
• A few smooth muscle fibres may be seen, but there is no muscular layer.
232 Practical Manual of Histology

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CERVIX

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify cervix under the light microscope.
2. Identify cervical glands.
3. Identify simple columnar epithelium of cervical canal and stratified squamous non-keratinized epithelium of
outer surface of cervix and the abrupt change of epithelium at external os.
4. Draw a diagram of microanatomy of cervix.

Function of cervical glands

Three points of identification

Practical Manual of Histology 233

Date:

Cervix (LS)
234 Practical Manual of Histology

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NOTES
Practical Manual of Histology 235

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FALLOPIAN TUBE/UTERINE TUBE

• Histologically there are three layers in the wall of fallopian tube—mucosa, muscular layer and serosa.
• The characteristic appearance of mucosa is its numerous folds projecting into the lumen. They appear to obscure
the lumen.
• Lining epithelium has ciliated simple columnar cells resting on basement membrane. Non-ciliated cells called
peg cells are also present.
• Lamina propria lies beneath the epithelium. It has loose connective tissue with fibroblast cells.
• There is no muscularis mucosa.
• Muscular layer has inner circular and outer longitudinal smooth muscle layers.
• Serosa is the outermost layer made up of mesothelium.

Answer the Following

1. What is the function of cilia in fallopian tube?

2. What is the function of clear or peg cells?

3. What is the role of muscular layer in fallopian tube?

236 Practical Manual of Histology

Date:

FALLOPIAN TUBE/UTERINE TUBE

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify fallopian tube under the light microscope.
2. Identify mucosal folds projecting in the lumen.
3. Identify simple columnar ciliated epithelium along with non-ciliated cells in the epithelium.
4. Identify lamina propria.
5. Identify muscular layer—inner circular and outer longitudinal.
6. Draw a diagram of microanatomy of fallopian tube/uterine tube/oviduct.

Function of cilia in uterine tube

Three points of identification

Practical Manual of Histology 237

Date:

Fallopian tube/uterine tube (TS)

238 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 239

Date:

PLACENTA
Placenta is the organ which connects the foetus to the mother. It is disc-shaped with a convex maternal and a
concave smooth foetal surface. Foetal surface is lined by chorionic plate and maternal surface exhibits cotyledons
due to placental septa.
It is better to study development of placenta before studying the microscopic structure. Presuming that it has
been studied we proceed with the details of microscopic anatomy.
There are three stages in the development of villi. They are as follows:
1. Primary villus: Central core is of cytotrophoblast which is surrounded by syncytiotrophoblast.
2. Secondary villus: Mesoderm is at the core, which is then surrounded by cytotrophoblast and then
syncytiotrophoblast.
3. Tertiary villus: Blood vessels enter the central core of mesoderm. So there is loose connective tissue with
foetal capillaries, and phagocytic cells called Hofbauer cells. This is then surrounded by cytotrophoblast,
and syncytiotrophoblast.
• In a section of placenta we can see the decidua basalis towards the maternal side. It has uterine glands, blood
vessels and some smooth muscle fibres.
• The foetal side has amnion and chorionic plate.
• In between are the chorionic villi—anchoring stem villi, primary villi, secondary villi and tertiary villi.
• Cytotrophoblast cells take up light basophilic stain and syncytiotrophoblast stains strongly basophilic on
H & E staining.
• Placental barrier can be seen comprising of:
o Syncytiotrophoblast cells
o Basement membrane of cytotrophoblast cells
o Basement membrane of foetal capillaries
o Endothelial cells.
240 Practical Manual of Histology

Date:

PLACENTA

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify placenta under the light microscope.
2. Identify stem, primary, secondary, tertiary villi, as the case may be.
3. Identify cytotrophoblast, syncytiotrophoblast and mesodermal core.
4. Identify foetal capillaries in villi and maternal RBCs in between villi.
5. Identify the placental barrier.
6. Draw the diagram of microanatomy of placenta.

Function of chorionic villi

Three points of identification

Practical Manual of Histology 241

Date:

Placenta
242 Practical Manual of Histology

Date:

UMBILICAL CORD

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify umbilical cord under the light microscope.
2. Identify two umbilical arteries and one umbilical vein.
3. Identify the Wharton’s jelly which has fibroblasts, collagen fibres and ground substance which form the
myxomatous tissue.
4. Identify the outer covering squamous epithelial cells of amnion.
5. Draw a diagram of microanatomy of umbilical cord.

Function of umbilical arteries and vein

Three points of identification

Practical Manual of Histology 243

Date:

Umbilical cord (TS)

244 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 245

Date:

CORPUS LUTEUM

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 52.3 Describe and identify the K/S SH Y Lecture, practical Written/skill
microanatomical features of assessment
cardioesophageal junction,
corpus luteum

• Graffian follicle ruptures releasing the oocyte. The fluid in antrum is also released. The follicle collapses and
later the granulosa cells and theca interna cells form corpus luteum.
• Granulosa cells and theca interna cells increase in size to form granulosa luteal cells and theca lutein cells
respectively.
• If menstruation occurs, then it forms corpus luteum of menstruation and if fertilisation occurs, then it is known
as corpus luteum of pregnancy.

Answer the Following

1. How is corpus luteum formed?

2. Which hormones are secreted by corpus luteum?

3. What happens to corpus luteum if fertilization fails to occur?

4. What happens to corpus luteum if fertilisation occurs?

246 Practical Manual of Histology

Date:

CORPUS LUTEUM

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify corpus luteum under the light microscope.
2. Identify granulosa luteal cells.
3. Identify theca lutein cells.
4. Make a diagram of the microanatomy of corpus luteum.

Function of theca lutein cells

Three points of identification

Practical Manual of Histology 247

Date:

Corpus luteum
248 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 249

Date:

BREAST

Number Competency Domain Level Core Teaching/learning Assessment Number required

Mammary gland of lactation

256 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 257

Date:

ENDOCRINE GLANDS

Practical Manual of Histology 263

Date:

Parathyroid gland
264 Practical Manual of Histology

Date:

PINEAL GLAND

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 43.3 Identify, describe and draw K/S SH N Lecture, practical Written/skill
microanatomy of olfactory assessment
epithelium, eyelid, lip, sclera-
corneal junction, optic nerve,
cochlea-organ of Corti, pineal gland

• A section shows calcified masses called corpora arenacea or brain sand. Corpora arenacea stains dark blue
with H & E staining.
• The pineal gland is covered with pia mater.
• 95% of the cells present in it are pinealocytes. They are polyhedral with large oval nucleus. Neuroglial cells,
mainly astrocytes lie in between the pinealocytes.
SPECIFIC LEARNING OBJECTIVES
At the end of the session the student should be able to:
1. Identify pineal gland under the light microscope.
2. Identify corpora arenacea.
3. Identify pinealocytes and neuroglial cells.
4. Draw a diagram of microanatomy of pineal gland.

Function of pinealocytes

Three points of identification

Practical Manual of Histology 265

Date:

Pineal gland
266 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 267

Date:

SUPRARENAL GLAND

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 52.1 Describe and identify the K/S SH Y Lecture, practical Written/skill
microanatomical features of assessment
gastro-intestinal system:
Oesophagus, fundus of stomach,
pylorus of stomach, duodenum,
jejunum, ileum, large intestine,
appendix, liver, gall bladder,
pancreas and suprarenal gland

• Suprarenal gland has outer cortex (develops from coelomic epithelium, i.e. mesoderm) and inner medulla
(develops from neural crest cells, i.e. ectoderm).
• Outermost covering capsule has capillaries.
• Cortex has three zones: Outermost—zona glomerulosa, middle—zona fasciculata and innermost—zona
reticularis. Capillaries are also present.
• Under the light microscope zona glomerulosa shows loops of basophilic columnar cells with dark spherical
nuclei and sinusoids in between the cell groups. Mineralocorticoids are secreted by these cells.
• Zona fasciculata has vertical columns of cells which appear light basophilic stained due to a large number of
vacuoles. They are known as spongiocytes. They secrete glucorticoids. Vertical sinusoids also run along the
columns of cells. Capillaries are also seen.
• Zona reticularis has a network of polygonal cells. They secrete sex steroid hormones. Capillaries can be seen in
between.
• Medulla has two types of cells—chromaffin cells (phaeochromocytes) (more numerous) and ganglion cells.
Medullary capillaries are also seen.
• Chromaffin cells are basophilic. The granules in the cytoplasm are precursors of epinephrine/adrenalin and
noradrenaline.
• Ganglion cells are interspersed between the chromaffin cells. They are larger than chromaffin cells and have
big nuclei with nucleoli.

Answer the Following

1. What are the three zones in cortex of suprarenal gland?

2. What is the embryological origin of cortex and medulla of suprarenal gland? Correlate it with type of secretions.
268 Practical Manual of Histology

Practical Manual of Histology 271

Date:

Spinal cord (TS)

272 Practical Manual of Histology

Date:

CEREBRUM

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify cerebrum under the light microscope.
2. Identify small, medium and large pyramidal cells.
3. Identify the six layers of cerebral cortex.
4. Draw a diagram of microanatomy of cerebrum.

Function of pyramidal cells

Three points of identification

Practical Manual of Histology 273

Date:

Cerebrum
274 Practical Manual of Histology

Date:

CEREBELLUM

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify cerebellum under the light microscope.
2. Identify folia of cerebellum.
3. Identify grey matter and white matter of cerebellum.
4. Identify the three layers of grey matter.
5. Identify Purkinje cells of middle layer.
6. Draw a labelled diagram of microstructure of cerebellum.

Function of Purkinje cells

Three points of identification

Practical Manual of Histology 275

Date:

Cerebellum
276 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 277

Date:

SPECIAL SENSES

Number Competency Domain Level Core Teaching/learning Assessment Number required

Date:

OPTIC NERVE

Number Competency Domain Level Core Teaching/learning Assessment Number required

(Y/N) method method to certify
AN 43.2 Identify, describe and draw K/S SH N Lecture, practical Written/skill
microanatomy of olfactory assessment
epithelium, eyelid, lip, sclero-
corneal junction, optic nerve,
cochlea-organ of Corti, pineal gland

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify optic nerve under the light microscope.
2. Identify bundles of axons of ganglion cells.
3. Identify central retinal vein and central retinal artery.
4. Draw a diagram of microanatomy of optic nerve.

Differentiate between peripheral and optic nerve

Three points of identification

Practical Manual of Histology 289

Date:

Optic nerve (TS)

290 Practical Manual of Histology

Date:

Function of oral mucosa

Three points of identification

Practical Manual of Histology 295

Date:

Lip
296 Practical Manual of Histology

Date:

NOTES
Practical Manual of Histology 297

Date:

COCHLEA AND ORGAN OF CORTI

What is bony labyrinth and membranous labyrinth?

• Labyrinth literally means a complex irregular network of passages. Bony labyrinth is a hollow meshwork of
cavities in the temporal bone in which lies the inner ear (vestibule, semicircular canals and cochlea).
• Membranous labyrinth is a collection of tubes (i.e. soft tissue) which are filled with fluid. They contain receptors
for the senses of balance and hearing.
What is cochlea?
It is a spiral tube of the inner ear cavity containing organ of Corti which produces nerve impulses in response to
sound vibrations. It appears like a shell of a snail. The tube spirals 2 3/4th times around a bony projection called
modiolus.
How is cochlea divided?
A shelf-like projection from the modiolus divides the cochlea incompletely into one tube above called scala vestibuli
and one tube below called scala tympani. Both these communicate at the apex through helicotrema. In the middle
is the organ of Corti which has another scala called scala media.
What is the organ of Corti and where is it found?
Organ of Corti is the receptor organ for hearing found in the cochlea between scala vestibuli and scala tympani.
What cells are found in the organ of Corti?
Single row of inner hair cells, 3 rows of outer hair cells, pillar cells, Deiter’s cells (also known as phalangeal cells),
Hensen’s cells, supporting cells.
What are the other structures found in the organ of Corti and cochlear duct?
• Reticular membrane—formed by apices of outer hair cells and phalangeal cells.
• Basilar membrane—extends from the tip of spiral lamina of modiolus to the outer wall of cochlea. It separates
cochlear duct from scala tympani.
• Tectorial membrane—it is gelatinous and produced by columnar cells on top of spiral limbus just medial to the
organ of Corti.
• Reissner’s membrane (vestibular membrane)—separates cochlear duct from scala vestibuli.
298 Practical Manual of Histology

Date:

COCHLEA AND ORGAN OF CORTI

SPECIFIC LEARNING OBJECTIVES

At the end of the session the student should be able to:
1. Identify cochlea and organ of Corti under the light microscope.
2. Identify scala vestibuli, scala tympani and scala media.
3. Identify Reissner’s membrane, tectorial membrane, basilar membrane and reticular membrane.
4. Identify inner and outer hair cells and tunnel of Corti.
5. Identify stria vascularis and limbus.
6. Draw a diagram of microanatomy of cochlea and organ of Corti.

Function of inner and outer hair cells of organ of Corti

Three points of identification

Practical Manual of Histology 299

Date:

Cochlea and organ of Corti

GUIDE TO STUDENTS FOR STAINING CHARACTERISTICS OF HISTOLOGY TISSUES
Characteristics of Haematoxylin and Eosin Staining in Various Tissues and Different Parts of Cells

Basophilic Violet (blue)

Acidophilic Lilac (pink)

Cell membrane Lilac (pink)

Nucleus Violet (blue)

Cytoplasm Lilac (pink)

Nucleoplasm Violet (blue)

Nucleoli Violet (blue)

Basement membrane Lilac (pink)

Elastic fibres Lilac (pink)

Collagen fibres Lilac (pink)

Reticular fibres Not stained by H & E

Elastic fibres of elastic cartilage Lilac (pink)

Hyaline cartilage Homogenous

Mucous Not stained by H & E

Serous inclusions Violet (blue)

Goblet cells Not stained by H & E

Muscle fibres Lilac (pink) (except their nucleus)

Bone matrix Lilac (pink)

Nerve fibre Lilac (pink)

Lacunae in cartilage Outline is violet

Lacunae in bones Outline is violet

Microvilli Lilac (pink) – deep staining

Keratin Lilac (pink)

Cilia Lilac (pink)

Stereocilia Lilac

Striated border Dark pink (lilac)

Corpora amylacea Eosinophilic (pink)

Lipid No staining by H & E

Corpora arenacea Basophilic (dark blue)

FEEDBACK FORM

1. Suggestions for improvement of diagrams

2. Suggestions for improvement of content.

3. Any other suggestions.

Note: Suggestion may be mailed at [email protected]; [email protected]

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