Diploma AHC questions (Histopathology)

1. Tissue must be placed in a suitable fixative as soon as it is removed from the body
to:
a. Prevent decomposition due to enzymatic activity
b. Permit various dehydrants to function properly
c. Prevent the rupture of cell membranes
d. Stabilise tissue carbohydrates
e. None of the above

2. The following fixative pairs are more likely to make solution acidic under certain
conditions?
a. Formalin & mercuric chloride
b. Formalin & osmium tetroxide
c. Mercuric chloride & osmium tetroxide
d. Potassium permanganate & glycoxal
e. All of the above

3. Dehydrating tissues in graded alcohols of increasing concentration is superior to

going directly into absolute alcohol because it will:
a. Cause less distortion of the tissue
b. Be less harmful to the tissue processor
c. Not harden tissue over a long period
d. Remove the fixative faster
e. None of the above

4. Decalcification occurs with all of the following methods EXCEPT:

a. Simple acid
b. Radiography
c. Chelation
d. Ion-exchange
e. None of the above

5. The process of saturating tissue with the medium that will be used for embedding is
called:
a. Clearing
b. Dehydration
c. Fixation
d. Infiltration
e. Embedding

6. Formalin pigment is generally formed in tissues in formaldehyde when the pH:

a. Rises above 6
b. Falls below 6
c. Is buffered to neutrality
d. All the above
e. None of the above

7. Alternate thick and thin zones within a section of very dense tissue, such as uterus
are most often caused by which of the following
a. Improper embedding
b. Inadequate processing
 c. Microtome or knife vibration
d. Wrong block temperature
e. None of the above

8. The components of Mayer’s haematoxylin are:

a. Haematoxylin, 95% alcohol, glycerol, ammonium alum, and acetic acid.
b. Haematoxylin, 95% alcohol, glycerol, and saturated ammonium alum
c. Haematoxylin, sodium iodate, chloral hydrate, and acetic acid.
d. Haematoxylin, sodium iodate, chloral hydrate, citric acid, and ammonium or
potassium alum.
e. None of the above

9. Differential staining of nuclei and cytoplasm with Geimsa solution is an example of:
a. Polychromasia
b. Metachromasia
c. Hyperchromasia
d. Metaplasia
e. None of the above

10. The selectivity for nuclear staining with Harris haematoxylin can be increased by
adding
a. Excess aluminium salts
b. Excess glycerol
c. Excess mercury
d. 50% alcohol
e. 100% alcohol

11. The process of saturating tissue with the medium that will be used for embedding is
called:
a. Clearing
b. Dehydration
c. Fixation
d. Infiltration
e. None of the above

12. Which of the following is not a constituent of 10% neutral buffered formalin?
a. Sodium phosphate trihydrate
b. Picric acid
c. Sodium phosphate
d. 37% - 40% formaldehyde
e. None of the above

13. Artifacts produced during tissue processing include

i. Folds
ii. Knife marks
iii. Debris
iv. Shrinkage
a. i, ii & iv
b. only iv
c. i, iii & iv
d. only iii
e. All of the above
 14. The following statements about neutral buffered 10% formalin are correct
i. The pH of the fixative is 8-12
ii. Methanol can be used to stabilize it
iii. Sodium chloride is added to maintain the right pH
iv. 37% formaldehyde solution is used

a. ii & iv
b. iii & iv
c. ii & iii
d. i & iv
e. All of the above

15. The following statements are TRUE about fixation

i. Acetic acid is both a coagulant and non-coagulant fixative

ii. Coagulant fixatives produce larger pore sizes in tissues than non-coagulant
iii. Paraffin wax penetrates non-fixed tissues more easily than fixed tissues
iv. Antibodies staining in fixed tissues is more feasible than non-fixed tissue

a. All of the above

b. i, ii & iv
c. ii, iii & iv
d. Only i
e. Only iv

16. Paraffin tissue blocks were left on ice for a longer duration. This will
i. Ensure temperature consistency of tissue and wax block
ii. Cause the tissue to swell
iii. Cause expansion and distortion of tissue section
iv. Enhance easy sectioning

a. All of the above

b. i, ii & iv
c. i & iv
d. ii & iii
e. Only iv

17. These are TRUE reason for manual tissue processing

i. Power failure
ii. Break down of tissue processor machine
iii. For whole brain
iv. Cheaper in cost

a. All of the above

b. i, ii & iii
c. i & ii
 d. iii & iv
e. Only iv

18. Special stains have these advantages over tinctorial stains

i. Substances stained can be quantified
ii. Controls are needed for special staining but not tinctorial stains
iii. Are used for diagnosis of cancers only
iv. Are relatively cheaper

a. i & ii
b. i, ii & iii
c. iii & iv
d. ii, iii & iv
e. All of the above

19. The importance of control preparations in histological staining includes

i. To determine if the stains work
ii. To assess the degree of non-specificity
iii. To determine if the reagent is still active
iv. To determine if the substance to stain is present

a. i, ii, iv
b. i, ii, iii
c. ii, iii & iv
d. None of the above
e. All of the above

20. The staining component of haematoxylin solution is

a. Haemalum
b. Hematein
c. Alum
d. Sodium iodate
e. Glycerol

21. Which of the special stains is specific for phosphate groups of DNA
i. Giemsa stain
ii. Feulgen stain
iii. Methyl green-pyronin
iv. Haematoxylin

a. i & iv
b. ii & iv
c. ii & iii
d. Only i
e. Only iv
22. A patient who had multiple blood transfusions is suspected to have haemolysis and a
liver biopsy is being sent to you. What special staining technique would you
recommend to confirm haemolysis?

a. Gordon and Sweet stain

b. Perl’s Stain
c. PAS
d. Masson Trichrome stain
e. Van Gieson

23. The purpose of embedding tissue in paraffin is to

a. Preserve antigenicity
b. Stabilize proteins
c. Provide support
d. Remove water from the cells
e. None of the above

24. To protect the exposed tissue during storage, paraffin blocks may need to be
a. Sealed
b. Filed
c. Stacked
d. Trimmed
e. None of the above

25. Alternate thick and thin zones within a section of very dense tissue, such as uterus, is
most often caused by which of the following?
a. Improper embedding
b. Inadequate processing
c. Microtome or knife vibration
d. Wrong block temperature
e. None of above

26. Components of living tissue do not require staining if they are to be examined by
a. Fluorescent microscopy
b. Light microscopy
c. Phase contrast microscopy
d. Transmission electron microscopy
e. All of the above

27. Which of the following underpins microwave - oven fixation of tissue?

a. Rapid molecular movement of fixative into tissue
b. Reaction between protein molecules
c. Ionising radiation
d. Rupture of hydrogen bonds
e. All of the above

28. To obtain a total magnification of 1000 with an ocular of 10x magnification, the
objective must have a magnification of:
a. 10x
b. 100x
c. 450x
d. 1000x
e. None of the above
29. To increase the amount of light when viewing a microscopic section, an adjustment
should be made in the:
a. Objective
b. Substage condenser
c. Numerical aperture
d. Iris diaphragm
e. None of the above

30. To prepare 50mL of 2% ferric chloride from a 10% solution, how many mL of the 10%
solution will be required?
a. 5
b. 10
c. 20
d. 25
e. 50

31. In a tertiary hospital pathology laboratory where the total number of cases received
annually is approximately 2500, large volumes of staining solutions are supposed to
be in stock. One of such solutions was prepared by dissolving 20g of solutes in 0.5L
of distilled water. What is the percentage of this solution?
a. 2%
b. 4%
c. 10%
d. 20%
e. 50%

32. In a histopathology laboratory a temperature of 45 - 50 oC is most commonly

associated with:
a. The floatation bath
b. The laboratory incubator
c. Paraffin dispenser
d. Hot air oven
e. Heating plate

33. 3. Determining the end - point of acid decalcification is very important because:
a. Calcium remaining in the tissues interferes with staining
b. Under-decalcification causes processing problems
c. Over-decalcification results in denaturation of cellular proteins
d. Prolonged acid treatment may compromise quality of nuclear staining.
e. None of the above

34. The clearing agent in the autoamated tissue processor was found to be cloudy and
turbid. It is most likely contaminated with:
a. Absolute alcohol
b. Bacteria
c. Water
d. Yeast
e. Oil

35. After completion of decalcification, the specimen should be:

a. Placed in acetone
b. Washed in 70% alcohol
c. Placed in fixative
d. Washed in water
e. Washed in xylene

36. For adequate clearing in xylene during routine processing, tissue should have a
maximum thickness of:
a. 1-2mm
b. 2-4mm
c. 4-6mm
d. 6-8mm
e. 8-10mm

37. Prolonged dehydration in the higher grades of alcohol will render the specimen:
a. Hard
b. Macerated
c. Porous
d. Toxic
e. Soft

38. The time needed for infiltration of paraffin wax is dependent upon all of the following
EXCEPT the:
a. Fixative used
b. Thickness of the specimen
c. Tissue type
d. Use of vacuum
e. None of the above

39. The choice of xylene as a clearing agent is because of its miscibility with:
a. Fixative and dehydrants
b. Fixatives and infiltrating media
c. Universal solvents
d. Dehydrants and infiltrating media
e. None of the above

40. The alcohols on the automated tissue processor should be changed on a regular
basis because:
a. The alcohols become saturated with bile from gall bladder specimens, which can be
absorbed by other tissues
b. Continuous transfer of formalin - fixed tissue into the alcohols cans create a
potentially explosive situation
c. Continuous transfer of formalin - fixed tissue into the alcohols can result in absorption
of moisture, which dilute the alcohols.
d. Alcohols become contaminated with fungal growth over prolonged usage.
e. None of the above

41. These must be recommended during histological sample reception

i. Tissues in a wrong fixative can be received and later transferred to the
appropriate fixative
ii. The patient data on the request form should match that on the specimen
container
iii. It is appropriate to change the identification number assigned to histological
cases during different stages of tissue processing
iv. Formalin kills all infectious agents thus there is no risk/hazard associated with
sample reception
a. i & ii
b. iii & iv
c. only ii
d. only iv
e. all of the above

42. Labeling of solutions is extremely important in managing chemical hazard. The

minimum information when labeling solutions therefore must be
i. Chemical name
ii. Hazards warnings and safety precautions
iii. Use/purpose of the solutions
iv. Expiration date

a. All of the above

b. ii, iii & iv
c. i, ii & iv
d. iii & iv
e. i & ii

43. You noticed a gallon of HCl spill over in the lab one early morning. There was nobody
there however. The precautionary measures you would take include
i. Place a hazard signboard and wait for the cleaner to clean it
ii. Add a neutralizing solution
iii. Ignore it and continue to work
iv. Record the incident in an accident book

a. None of the above

b. i & iv
c. i & iii
d. ii & iv
e. Only ii

44. A xylene reagent used in tissue processing could clear again because it had lost its
strength. It can be
a. Disposed off in the drainage
b. Bottled for recycling
c. Left in the sun to evaporate
d. Mixed with more water and disposed off in the drainage
e. None of the above

45. While using a 10x objective to view an H&E slide stained section, the section
appears bright with well-stained nuclei. When the objective is changed to 40x, the
section appears dark and detail is obscured. This most likely will be corrected by:
a. Using the 90x objective
b. Cleaning the eye piece
c. Restaining and decreasing the haematoxylin
d. Slightly opening the iris diaphragm
e. None of the above

46. To maximise the resolving power of a light microscope, the substage condenser
should be adjusted to sharply focus the image of the:
a. Sub stage diaphragm
b. Lamp filament
c. Specimen slide
d. Field diaphragm
e. Any of the above

47. Which of the following is not crucial for retrospective studies in future researches.
a. Proper filing and storage of stained slides
b. Proper filing and storage of tissue blocks
c. Reports filing and database
d. Specimen preservation and potting
e. All of the above

48. Recent erratic power supply resulted in the failure of completion of overnight-
automated tissue processing, causing the basket loaded with tissue cassettes to stay
in a xylene compartment for a protracted period of time without progressing to the
stage of impregnation. Which of the following is a probable effect on the tissues and
the line of action for remedy.
a. Tissue shrinkage; rehydrate and return to dehydration before impregnation
b. Hardening of tissues; manually reduce impregnation time
c. Softening of tissue; manually increase the impregnation time
d. Tissue becomes brittle: add phenol to the xylene
e. All of the above

49. While performing the daily temperature check of the embedding centre paraffin
reservoir, it is noted that the temperature is 12 oC above the melting point of paraffin.
The most appropriate action is to first:
a. Drain and replace the paraffin
b. Check the thermostat setting
c. Recheck the temperature in 24hrs.
d. Switch off the embedding centre for the temperature to stabilise.
e. None of the above

50. Microscopic review of formalin – fixed tissue section demonstrated fine brown to
black artifactual pigment. This would have been prevented if the formalin solution had
been made:
a. Alkaline
b. Neutral
c. Acidic
d. Amphoteric
e. none of the above
Key
1 A
2 A
3 A
4 B
5 D
6 A
7 C
8 D
9 A
10 A
11 D
12 A
13 E
14 A
15 B
16 A
17 B
18 A
19 B
20 A
21 D
22 B
23 C
24 A
25 C
26 D
27 A
28 B
29 C
30 B
31 B
32 A
33 D
34 C
35 D
36 B
37 A
38 A
39 D
40 C
41 C
42 C
43 D
44 B
45 D
46 A
47 D
48 B
49 B
50 B