Pathology (PHARM:514)
Practical –1
DR. DALAQ AIYSHA | ASSISTANT PROFESSOR
Division
Study of pathological slides
Examination of different
body fluids
Tests for various specimens
of clinical importance
Activity 1
When you hear the word
histopathology, what
comes to mind?
Lecture:1
HISTOPATHOLOGY
Learning Objectives
Understand the importance of histopathology in
disease diagnosis and therapy
Learn basic tools and techniques used in a
histopathology lab
Study the principle and purpose of common staining
methods, i.e., H&E
Recognize how staining helps in visualizing tissues and
identifying pathological changes
Definition
A branch of pathology
Histopathology is the microscopic study of
diseased tissues.
It involves examining thin tissue sections under a
microscope after they have been fixed,
processed, embedded, sectioned, and stained.
What is Histopathology?
“Histo” = tissue, “Pathology” = study of disease
Uses microscopy to identify abnormal structures in tissues
Helps in the diagnosis of diseases by comparing normal
vs diseased tissue
Relies heavily on the Hematoxylin & Eosin (H&E) stain
Importance
Medicine
Confirms diagnoses of diseases like cancer, infections,
inflammatory diseases
Pharmacy
Detects drug-induced tissue damage (e.g.,
hepatotoxicity, nephrotoxicity)
Helps in drug development by studying tissue response
Assists in identifying adverse drug reactions (ADRs).
Lab Safety Rules
Always wear PPE: lab coat, gloves, and safety goggles
Handle chemicals (formalin, xylene) only in a fume hood
Treat all specimens as potentially infectious – follow biohazard
protocols.
Operate equipment safely – use microtome blade guards and
caution
Keep workbench clean and store chemicals properly.
Report accidents or faulty equipment immediately to
supervisor (broken slides/microscopes)
Processes in Histopath
Lab
1. Specimen Collection & Reception – tissue received in
fixative (usually formalin) with proper labeling.
2. Gross Examination (Grossing) – description, measurement,
and trimming of tissue for processing.
3. Fixation – preservation of tissue architecture (commonly 10%
formalin).
4. Tissue Processing – dehydration, clearing, and paraffin
infiltration using tissue processor.
5. Embedding – tissue placed into paraffin wax blocks.
6. Sectioning (Microtomy) – thin slices (3–5 µm) cut with a
microtome.
Processes in Histopath
Lab
1. Floating & Mounting – sections floated on a warm
water bath, then mounted on glass slides.
2. Staining – routine Hematoxylin & Eosin (H&E), plus
special/histochemical stains as needed.
3. Cover slipping – permanent mounting with cover
glass for preservation.
4. Microscopic Examination – slides examined by
pathologists for diagnosis.
5. Reporting – findings documented in the pathology
report.
6. Archiving – storage of tissue blocks and slides for
future reference.
Essential Tools in
Histopathology
1. Tissue Processing 3. Staining
• Forceps & Scalpels – tissue handling and • Staining racks/jars – hold slides during staining
cutting
• Automated Stainer – routine H&E and special
• Tissue cassettes – hold specimens during stains
processing
• Reagent bottles & pipettes – for dyes and
• Tissue processor – dehydration, clearing, solutions
infiltration
4. Microscopy & Imaging
• Paraffin wax dispenser & embedding
station – block preparation • Light microscope (compound) – routine
diagnosis
2. Sectioning
• Multiheaded microscope – teaching & group
• Microtome – cuts thin tissue sections (3–5 diagnosis
µm)
• Fluorescence microscope – immunostains
• Cryostat – frozen section cutting for rapid
diagnosis • Digital slide scanner – whole-slide imaging &
archiving
• Water bath – stretches tissue sections
5. General Lab Tools
before mounting
• Slide warmers / hot plates – dry and fix • Glass slides & cover slips – for mounting sections
sections • Fume hood – handling formalin and xylene safely
Microscope
Types – Light (routine), Phase-contrast (live cells), Polarizing (crystals),
Fluorescence (special stains), Electron (ultrastructure).
Parts – Optical (eyepiece, objectives), Illumination (lamp, condenser,
diaphragm), Mechanical (stage, knobs, base).
Magnification – Eyepiece × objective (4x, 10x, 40x, 100x oil immersion).
Handling – Carry properly, clean with lens paper, focus under low power
first, avoid coarse focus on high/oil immersion.
Oil Immersion – Drop oil, use 100x objective, clean after use.
Care – Keep dust-free, switch off light, cover microscope, store with the
lowest objective in place.
Parts of a Micorscope
Activity 2
Staining Purpose
Staining Methods
Routine Staining
• H&E (Hematoxylin & Eosin): Nuclei blue-purple, cytoplasm pink
• Most commonly used; shows general tissue architecture
Special Stains
• PAS: Highlights carbohydrates & glycogen (magenta) – fungi,
basement membranes
• Masson’s Trichrome: Differentiates collagen (blue/green) &
muscle (red) – fibrosis
• Reticulin (Silver stain): Shows reticular fibers – liver & marrow
architecture
• Ziehl-Neelsen: Detects acid-fast bacteria (red) – tuberculosis
• Giemsa: Blood cells & parasites – malaria, hematology
Staining Methods
Immunohistochemistry (IHC)
• Uses antibodies to detect specific proteins
• Guides targeted drug therapy
Fluorescent Stains
• Fluorochromes visualize structures under a
fluorescence microscope
• Example: DAPI stains nuclei blue
Activity 3
How do we choose which
staining method to use?
Summary
Study of diseased tissues at the cellular level.
Uses microscopes and tissue preparation techniques
H&E and special stains reveal tissue structures and
pathology.
IHC and fluorescent stains detect specific proteins and
components.
Guides diagnosis, therapy, and research.
Enhances understanding of disease mechanisms and
clinical decisions
Homework
Homework Assignment:
Exploring Histopathology
Stains Online
Thank You
Questions?
Lab Record Book
Practical Copy Guidelines
Each student must maintain a practical copy for
histopathology.
Today’s lecture content (intro + staining methods) should be
written as your first entry.
Diagrams (hand-drawn or printed) are encouraged for stains and
tissue slides.
SUBMISSION BEFORE THE
START OF THE NEXT
LECTURE
Checked regularly and counts towards the assessment.
Format
Copy should include:
Index page (at the start of the copy)
Title of practical along with date
Objective
Principle
Materials
Procedure
Observations (drawings/images/notes)
Conclusion
