General Histopathology

1. Which of the following fixatives is most commonly used in histopathology?

A. Glutaraldehyde
B. Ethanol
C. Formalin
D. Acetone
2. The main purpose of tissue fixation is to:
A. Make tissues soft
B. Remove water
C. Preserve tissue structure
D. Improve staining
3. Which of the following is a common embedding medium in histopathology?
A. Wax
B. Paraffin
C. Agar
D. Gelatin
4. Hematoxylin stains which cellular component?
A. Cytoplasm
B. Lipids
C. Nucleus
D. Mitochondria
5. What is the function of eosin in H&E staining?
A. Stains nucleus blue
B. Stains connective tissue green
C. Stains cytoplasm pink
D. Fixes tissue

Special Stains & Techniques

6. Which stain is used to detect acid-fast bacilli (e.g., Mycobacterium tuberculosis)?

A. PAS
B. Ziehl-Neelsen
C. Congo red
D. Silver stain
7. Periodic acid–Schiff (PAS) stain is used to demonstrate:
A. Glycogen and mucopolysaccharides
B. Lipids
C. Fungi only
D. Amyloid
8. What type of microscopy is used for viewing histopathological slides?
A. Electron microscope
B. Fluorescence microscope
C. Light microscope
 D. Phase-contrast microscope

Tissue Processing and Microtomy

9. The instrument used to cut thin sections of paraffin-embedded tissue is called:

A. Cryostat
B. Microtome
C. Centrifuge
D. Autoclave
10. During dehydration in tissue processing, which reagent is commonly used?
A. Formalin
B. Water
C. Xylene
D. Alcohol

Clinical Applications and Interpretation

11. Granulomas are commonly seen in:

A. Hepatocellular carcinoma
B. Tuberculosis
C. Diabetes mellitus
D. Lymphoma
12. Which of the following conditions is associated with amyloid deposition?
A. Multiple myeloma
B. HIV
C. Cystic fibrosis
D. Influenza
13. In histopathology, malignancy is best diagnosed based on:
A. Nuclear morphology
B. Cytoplasmic color
C. Tissue thickness
D. Fixative used
14. Which histological feature indicates a chronic inflammatory response?
A. Neutrophil infiltration
B. Lymphocyte and plasma cell infiltration
C. Edema
D. Hyperemia
15. What is the hallmark of coagulative necrosis under the microscope?
A. Liquefied tissue
B. Preserved tissue architecture
 C. Fat globules
D. Granuloma formation

Case-Based MCQ Questions in Histopathology

Case 1: Granulomatous Inflammation

Explanation: A fibroadenoma is a benign breast tumor commonly seen in young women, with classic
features as described.

Histopathology Technical MCQs

1. Which of the following fixatives is most commonly used in routine histopathology?

A. Carnoy's solution
B. Bouin’s solution
C. 10% neutral buffered formalin
D. Glutaraldehyde
Answer: C

2. What is the main purpose of tissue fixation?

A. To stain the tissue
B. To dehydrate the tissue
C. To preserve tissue structure and prevent autolysis
D. To embed the tissue in paraffin
Answer: C

3. Which of the following is the correct sequence in routine tissue processing?

A. Fixation → Clearing → Dehydration → Embedding
B. Fixation → Dehydration → Clearing → Embedding
C. Fixation → Embedding → Dehydration → Clearing
D. Embedding → Fixation → Dehydration → Clearing
Answer: B

4. Which reagent is commonly used for the clearing step during tissue processing?
A. Ethanol
B. Formalin
C. Xylene
D. Paraffin wax
Answer: C
5. What is the ideal thickness of tissue sections cut on a microtome for routine H&E
staining?
A. 1–2 µm
B. 3–5 µm
C. 8–10 µm
D. 15–20 µm
Answer: B

6. What is the main staining method used in routine histopathology?

A. Ziehl-Neelsen
B. Giemsa
C. Hematoxylin and Eosin (H&E)
D. PAS
Answer: C

7. Hematoxylin stains which cellular structure?

A. Cytoplasm
B. Nuclei
C. Mitochondria
D. Cell membrane
Answer: B

8. What is the purpose of using eosin in H&E staining?

A. To stain the nuclei
B. To stain the background
C. To stain the cytoplasm and extracellular matrix
D. To fix the tissue
Answer: C

9. Which microtome is most commonly used in routine histopathology labs?

A. Rotary microtome
B. Sliding microtome
C. Ultramicrotome
D. Cryostat
Answer: A
10. Which of the following is a common cause of tissue folding in sections?
A. Over-fixation
B. Dull microtome blade
C. Excess dehydration
D. Poor staining technique
Answer: B

11. Which substance is used for embedding tissues in routine histopathology?

A. Paraffin wax
B. Agar
C. Gelatin
D. Resin
Answer: A

12. What is the main function of alcohol in tissue processing?

A. Clearing
B. Dehydration
C. Fixation
D. Staining
Answer: B

13. What is the likely outcome if tissue is not properly fixed before processing?
A. Enhanced staining
B. Better sectioning
C. Tissue autolysis and poor morphology
D. Sharper nuclear details
Answer: C

14. Cryostat is used for:

A. Embedding
B. Cutting paraffin blocks
C. Cutting frozen sections
D. Fixation
Answer: C
15. A common cause of air bubbles in stained sections is:
A. Poor dehydration
B. Excessive clearing
C. Incomplete mounting
D. Overfixation
Answer: C

16. What is the recommended temperature for paraffin wax embedding?

A. 25°C
B. 37°C
C. 45°C
D. 55–60°C
Answer: D

17. Which fixative is best for preserving fine cellular details?

A. 10% formalin
B. Bouin’s solution
C. Glutaraldehyde
D. Osmium tetroxide
Answer: C

18. What happens if xylene is not completely removed before embedding?

A. Tissue becomes too hard
B. Paraffin wax does not infiltrate properly
C. Tissue over-fixes
D. Sectioning becomes easier
Answer: B

19. What component of hematoxylin acts as a mordant?

A. Acetic acid
B. Ferric chloride
C. Eosin
D. Alcohol
Answer: B
20. Which of the following is NOT a feature of well-fixed tissue?
A. No shrinkage
B. Preserved morphology
C. Prevents autolysis
D. Hard and brittle texture
Answer: D

Case Scenario:
A 35-year-old male from a rural area presents with a chronic cough, night sweats, and weight
loss. A biopsy from a lung lesion is submitted for histopathological examination.

Histology Findings:

 Granulomas composed of epithelioid cells, Langhans-type multinucleated giant cells.

 Central caseous necrosis.

Question:
What is the most likely diagnosis and what type of inflammatory response is seen?

Answer:
Diagnosis: Pulmonary tuberculosis.
Type of Inflammation: Chronic granulomatous inflammation with caseation necrosis.

Explanation:
The presence of granulomas with caseous necrosis is classic for TB. The body's immune
response attempts to contain the infection by forming granulomas. Langhans giant cells are
typical in TB.

✅ Case 2: Hepatocellular Carcinoma (HCC)

Case Scenario:
A 50-year-old male with a history of chronic hepatitis B presents with right upper quadrant pain
and weight loss. A liver biopsy is performed.

Histology Findings:

 Trabecular arrangement of atypical hepatocytes.

 Increased nuclear-to-cytoplasmic ratio, prominent nucleoli, and mitotic figures.
 Pseudoglandular structures.

Question:
What is the most likely diagnosis?
Answer:
Hepatocellular carcinoma (HCC).

Explanation:
HCC arises in the background of chronic liver disease (like HBV infection). Histology shows
atypical hepatocytes in thickened trabeculae and pseudoglandular structures.

✅ Case 3: Squamous Cell Carcinoma of the Cervix

Case Scenario:
A 42-year-old woman presents with abnormal vaginal bleeding. A cervical biopsy is done.

Histology Findings:

 Invasive nests of malignant squamous cells.

 Keratin pearls and intercellular bridges.
 Desmoplastic stroma.

Question:
What is the diagnosis and its possible etiology?

Answer:
Diagnosis: Squamous cell carcinoma of the cervix.
Etiology: Persistent infection with high-risk Human Papillomavirus (HPV), especially types 16
and 18.

Explanation:
Histopathology shows keratinizing malignant squamous cells. HPV plays a major role in
carcinogenesis through E6 and E7 oncoproteins, which inhibit p53 and Rb.

✅ Case 4: Acute Appendicitis

Case Scenario:
A 19-year-old male presents with right lower quadrant abdominal pain, fever, and leukocytosis.
The appendix is removed.

Histology Findings:

 Neutrophilic infiltration involving all layers of the appendix wall (mucosa to serosa).
 Edema and congestion.
Question:
What is the diagnosis?

Answer:
Acute appendicitis.

Explanation:
The hallmark of acute appendicitis on histology is transmural neutrophilic infiltration. If not
treated, it can lead to perforation and peritonitis.

✅ Case 5: Schistosomiasis (Liver)

Case Scenario:
A 30-year-old farmer from the Oromia region presents with hepatosplenomegaly and signs of
portal hypertension.

Histology Findings:

 Granulomatous reaction around schistosome eggs.

 Periportal fibrosis ("pipestem fibrosis").

Question:
What is the likely diagnosis and causative agent?

Answer:
Diagnosis: Hepatic schistosomiasis.
Causative Agent: Schistosoma mansoni (most common in Ethiopia).

Explanation:
The immune response to trapped eggs leads to granulomas and fibrosis. Chronic infection leads
to portal hypertension even in the absence of cirrhosis.

✅ Case 6: Hydatid Disease (Liver)

Case Scenario:
A 28-year-old female from a pastoralist community presents with right upper quadrant pain.
Imaging shows a cystic liver lesion. Surgical resection is performed.

Histology Findings:

 Laminated acellular cyst wall.

 Inner germinal layer.
  Presence of scolices and hooklets.

Question:
What is the most likely diagnosis and causative organism?

Answer:
Diagnosis: Hydatid cyst of the liver.
Causative Agent: Echinococcus granulosus.

Explanation:
The cyst wall and presence of scolices are pathognomonic. It’s a zoonotic disease prevalent in
rural Ethiopia, often involving sheep and dogs.

Technical questions
1. What is the purpose of fixation in histopathology?
2. Why is 10% neutral buffered formalin preferred over other fixatives?
3. List the steps of routine tissue processing in histopathology.
4. What is the purpose of clearing in tissue processing?
5. Why is paraffin wax used in embedding?
6. What is the optimal thickness of tissue sections cut for routine histology?
7. What is the purpose of hematoxylin and eosin (H&E) staining?
8. What are common causes of poor tissue fixation?
9. Why is microtomy important in histopathology?
10. List common histopathology artifacts and their causes
11.What is decalcification and when is it necessary?
12.What are safety measures in a histopathology lab?
 1. Answer:
To preserve tissues in as life-like a state as possible and prevent autolysis and
putrefaction.

Explanation:
Fixatives like formalin cross-link proteins and stabilize cellular structures. In Ethiopia, 10%
neutral buffered formalin is commonly used due to availability and cost-effectiveness.

2. Answer:

Because it is cheap, effective, and preserves tissue morphology without excessive hardening.

Explanation:
Buffered formalin maintains pH stability and reduces artifacts that might appear with unbuffered
or acidic fixatives.

3. Answer:

1. Fixation
2. Dehydration
3. Clearing
4. Infiltration (embedding with paraffin)
5. Embedding
6. Sectioning
7. Staining
8. Mounting

Explanation:
Each step is critical for preparing tissue for microscopic examination. For instance, dehydration
removes water using increasing concentrations of alcohol.
4. Answer:
To remove alcohol and prepare the tissue for paraffin infiltration.

Explanation:
Clearing agents like xylene or substitutes make tissues translucent and miscible with paraffin
wax.

5, Answer:
Because it provides a supportive medium to section the tissue thinly for microscopy.

Explanation:
Paraffin is solid at room temperature but can be melted for infiltration and embedding. It holds
the tissue in place during sectioning.

6. Answer:
3 to 5 micrometers.

Explanation:
This thickness ensures enough structural detail without being too thick to allow light
transmission under the microscope.

7. Answer:
To differentiate and visualize tissue components.

Explanation:
Hematoxylin stains nuclei blue/purple, and eosin stains cytoplasm and extracellular matrix pink.
It’s the most common routine stain.
8. Answer:

 Large tissue size

 Inadequate fixative volume
 Delayed fixation
 Poor fixative penetration

Explanation:
Tissues should be no more than 5 mm thick and placed in at least 10–20 times their volume of
fixative immediately after removal.

9. Answer:
It produces thin tissue sections suitable for microscopic examination.

Explanation:
A rotary microtome is commonly used. Blunt blades, improper wax temperature, or poor
trimming can cause artifacts like chatter or wrinkles.

10. Answer:

 Chatter: vibration during sectioning

 Folding: improper section placement
 Cracking: over-dehydration
 Staining artifacts: due to improper reagent handling

Explanation:
Recognition and avoidance of artifacts are crucial for accurate diagnosis.

11. Answer:
Removal of calcium from tissues like bone to allow sectioning.

Explanation:
Acids (e.g., formic acid) or chelating agents (e.g., EDTA) are used before processing to soften
hard tissues.
12. Answer:

 Use PPE (gloves, lab coat, mask)

 Proper ventilation when handling xylene/formalin
 Labeling chemicals correctly
 Proper disposal of sharps and biohazards

Explanation:
Chemical exposure and biohazard risks are high in pathology labs, especially with common
reagents like formalin and xylene.
A 30-year-old male from rural Ethiopia presents with a chronic cough, weight loss, and night sweats. A
lymph node biopsy is performed, and histopathological examination shows caseating granulomas with
Langhans giant cells.

Q1. What is the most likely diagnosis?

A. Sarcoidosis

B. Tuberculosis

C. Leprosy

D. Histoplasmosis

Answer: B. Tuberculosis

Explanation: Caseating granulomas with Langhans giant cells are typical in tuberculosis, which is
endemic in Ethiopia.

Case 2: Cervical Cancer Screening

A 45-year-old woman attends a cervical cancer screening clinic. A Pap smear shows dysplastic squamous
epithelial cells. A biopsy of the cervix is taken and shows neoplastic squamous cells invading the stroma.

Q2. What is the most likely histopathological diagnosis?

A. Cervical intraepithelial neoplasia (CIN)

B. Squamous cell carcinoma

C. Adenocarcinoma of the cervix

D. Chronic cervicitis

Answer: B. Squamous cell carcinoma

Explanation: Invasion of the stroma by dysplastic squamous cells indicates squamous cell carcinoma.

Case 3: Liver Pathology

A 50-year-old male with a history of chronic alcohol consumption presents with abdominal swelling and
jaundice. A liver biopsy shows ballooning degeneration of hepatocytes, Mallory bodies, and fibrosis.

Q3. What is the most likely histopathological diagnosis?

A. Hepatocellular carcinoma

B. Cirrhosis due to hepatitis B

C. Alcoholic hepatitis

D. Fatty liver disease

Answer: C. Alcoholic hepatitis

Explanation: Mallory bodies and ballooning degeneration are classic features of alcoholic hepatitis.

Case 4: Schistosomiasis

A 22-year-old university student from the Awash River basin presents with hematuria. A bladder biopsy
reveals granulomatous inflammation and calcified ova with terminal spines.

Q4. What is the causative organism?

A. Schistosoma haematobium

B. Schistosoma mansoni

C. Echinococcus granulosus

D. Trichuris trichiura
Answer: A. Schistosoma haematobium

Explanation: Terminal spined ova are characteristic of S. haematobium, commonly affecting the bladder
in Ethiopia.

Case 5: Breast Lump

A 35-year-old woman presents with a painless, mobile breast lump. Histopathology reveals ducts lined
by two layers of epithelium and fibrous stroma with no atypia.

Q5. What is the most likely diagnosis?

A. Fibroadenoma

B. Fibrocystic change

C. Invasive ductal carcinoma

D. Ductal carcinoma in situ

Answer: A. Fibroadenoma

Case 1: Tuberculosis Lymphadenitis

Case:
A 25-year-old woman presents with a painless swelling in her neck that has gradually increased
over the past month. She has low-grade fever and night sweats. A fine needle aspiration of the
lymph node is sent for histopathologic examination.

Histopathology Findings:
Granulomatous inflammation with central caseous necrosis
Questions:

1. What is the most likely diagnosis?

2. What special stain can be used to confirm the causative agent?

Case 2: Cervical Carcinoma

Case:
A 45-year-old woman complains of post-coital bleeding and foul-smelling vaginal discharge. A
biopsy of the cervix is taken.

Histopathology Findings:
Disordered epithelial growth with pleomorphic cells, increased mitotic activity, and invasion into
the underlying stroma.

Questions:

1. What is the most likely diagnosis?

2. Which virus is most commonly associated with this condition?
3. Mention two histological features that indicate malignancy.
4. How is this cancer graded histologically?
5. Why is early histopathological diagnosis important in cervical carcinoma?

Case 3: Liver Schistosomiasis

Case:
A 35-year-old farmer from the Awash River Valley presents with abdominal distention and
hepatosplenomegaly. A liver biopsy is performed.

Histopathology Findings:
Granulomas surrounding Schistosoma eggs, fibrosis, and portal tract thickening.

Questions:

1. What is the diagnosis?

2. Which species of Schistosoma is most likely involved?
3. Describe the granulomatous response seen in schistosomiasis.
4. How does the histology differ from viral hepatitis?
5. What public health measures can reduce the incidence of this condition?
Case 4: Kaposi Sarcoma (HIV-related)

Case:
A 30-year-old HIV-positive man presents with multiple purple skin nodules. A biopsy of a skin
lesion is taken.

Histopathology Findings:
Spindle-shaped cells, slit-like vascular spaces, and hemosiderin-laden macrophages.

Questions:

1. What is the most likely diagnosis?

2. Which virus is associated with this tumor?
3. List the three histologic phases of Kaposi sarcoma.
4. Why is histology important in differentiating Kaposi sarcoma from other vascular
tumors?
5. How does immunosuppression affect the progression of this disease?

Case 5: Chronic Pyelonephritis

Case:
A 50-year-old man with a history of recurrent urinary tract infections presents with flank pain. A
nephrectomy specimen is submitted for histopathology.

Histopathology Findings:
Thyroidization of renal tubules, interstitial fibrosis, chronic inflammatory infiltrate, and
glomerulosclerosis.

Questions:

1. What is the diagnosis?

2. Describe the term “thyroidization” in renal histology.
3. How does chronic pyelonephritis differ from acute pyelonephritis histologically?
4. What are common causes of chronic pyelonephritis in Ethiopia?
5. What complications can arise from this conditio