Histopathology Technologist Prometric exam questions with answers

1. Question 1Immunohistochemistry and Molecular Techniquesmedium

   Which chromogen gives a brown reaction product with horseradish peroxidase in IHC?

   1. AFast Red, which gives a red product and is normally used with alkaline phosphatase
   2. BBCIP/NBT, which gives a blue-purple product with alkaline phosphatase detection
   3. CDAB (3,3'-diaminobenzidine), which gives a brown product with peroxidase
   4. DEosin Y, which is a pink counterstain and not an enzyme chromogen
   Show answer and explanation▾

   Answer: C. DAB (3,3'-diaminobenzidine), which gives a brown product with peroxidase

   DAB reacts with HRP and hydrogen peroxide to make an insoluble brown product. Fast Red and BCIP/NBT are used with alkaline phosphatase, and eosin is a counterstain.

   Reference: Agilent Dako, Education Guide: Immunohistochemical Staining Methods, 6th ed. (2013)

2. Question 2Immunohistochemistry and Molecular Techniqueshard

   For in situ hybridization or other molecular tests on FFPE tissue, which fixation practice most protects nucleic acids?

   1. AFixing in unbuffered formalin for several weeks
   2. BFixing in neutral buffered formalin for a controlled time, avoiding prolonged fixation and acid decalcifiers
   3. CUsing strong nitric acid to decalcify before testing
   4. DLeaving the tissue unfixed for 24 hours before processing
   Show answer and explanation▾

   Answer: B. Fixing in neutral buffered formalin for a controlled time, avoiding prolonged fixation and acid decalcifiers

   Controlled time in buffered formalin limits cross-linking and DNA fragmentation, while acid and long delays damage nucleic acids. Unbuffered formalin and delays degrade the target.

   Reference: Allison KH et al., Estrogen and Progesterone Receptor Testing in Breast Cancer: ASCO/CAP Guideline Update, J Clin Oncol 2020;38:1346-1366

3. Question 3Laboratory Safety and Professional Practicemedium

   How should xylene waste from the stainer be managed?

   1. APour it into the general sink with plenty of running water to dilute it
   2. BLet it evaporate in an open tray in the fume cupboard to reduce volume
   3. CMix it with formalin and pour the mixture into the laboratory drains
   4. DCollect in labeled, closed containers for hazardous waste disposal
   Show answer and explanation▾

   Answer: D. Collect in labeled, closed containers for hazardous waste disposal

   Xylene is flammable and toxic and must be collected in labeled containers for licensed disposal. Drains, open evaporation and mixing with formalin are unsafe.

   Reference: Suvarna SK, Layton C, Bancroft JD, Bancroft's Theory and Practice of Histological Techniques, 8th ed. (2019)

4. Question 4Microtomy and Frozen Sectionsmedium

   What temperature is typically used for cryostat cutting of most unfixed tissue for frozen section?

   1. AAbout 4 degrees Celsius
   2. BAbout 37 degrees Celsius
   3. CAbout -80 degrees Celsius in the cabinet
   4. DAbout -20 to -25 degrees Celsius
   Show answer and explanation▾

   Answer: D. About -20 to -25 degrees Celsius

   Most tissues cut best around -20 to -25 degrees, and fatty tissue needs colder. A refrigerator temperature or body temperature would not freeze the tissue, and the chamber is not run at -80.

   Reference: Carson FL, Hladik C, Histotechnology: A Self-Instructional Text, 5th ed. (2021)

5. Question 5Specimen Reception, Fixation and Grossing Supporteasy

   Which fixative is the routine choice for most surgical pathology specimens?

   1. A70 percent ethanol in water, which is the standard for all surgical specimens
   2. B10 percent neutral buffered formalin, which preserves tissue morphology
   3. CGlutaraldehyde in cacodylate buffer, which suits routine light microscopy best
   4. DCarnoy fluid with acetic acid, which preserves all antigens for routine work
   Show answer and explanation▾

   Answer: B. 10 percent neutral buffered formalin, which preserves tissue morphology

   Neutral buffered formalin (about 4 percent formaldehyde) penetrates well, preserves morphology and works with most stains and IHC. Ethanol and Carnoy fluid are used for special purposes, and glutaraldehyde is for electron microscopy.

   Reference: Suvarna SK, Layton C, Bancroft JD, Bancroft's Theory and Practice of Histological Techniques, 8th ed. (2019)

6. Question 6Quality Control and Troubleshootingmedium

   Sections show a blue-black granular precipitate on the tissue after H&E. What is the most probable cause?

   1. AUnfiltered hematoxylin with precipitated dye
   2. BUnder-fixation of the specimen, which gives a pale and patchy section instead
   3. CCutting the section too thick, which gives dense dark staining but no crystals
   4. DPoor clearing with xylene, which gives cloudy sections and wax remnants
   Show answer and explanation▾

   Answer: A. Unfiltered hematoxylin with precipitated dye

   Hematoxylin that has oxidized forms a metallic sheen and precipitates on the slide, so filtering is needed. Under-fixation, thickness and clearing produce different artifacts.

   Reference: Suvarna SK, Layton C, Bancroft JD, Bancroft's Theory and Practice of Histological Techniques, 8th ed. (2019)

7. Question 7Specimen Reception, Fixation and Grossing Supporthard

   Bone biopsies must be decalcified before sectioning. Which statement about the choice of decalcifier is correct?

   1. AConcentrated nitric acid is preferred because it preserves nuclear detail best
   2. BDecalcification is done before fixation to save time
   3. CEDTA is slow but preserves antigens and nucleic acids better than strong mineral acids
   4. DAcid decalcifiers do not need an endpoint check
   Show answer and explanation▾

   Answer: C. EDTA is slow but preserves antigens and nucleic acids better than strong mineral acids

   EDTA chelates calcium slowly and is gentler on antigens and DNA. Strong acids are fast but damage nuclear stain and molecular targets. Tissue is fixed first, and an end point test is needed to avoid over-decalcification.

   Reference: Suvarna SK, Layton C, Bancroft JD, Bancroft's Theory and Practice of Histological Techniques, 8th ed. (2019)

8. Question 8Tissue Processing and Embeddingeasy

   What is the correct order of the main steps of routine tissue processing?

   1. ADehydration, clearing, then paraffin infiltration
   2. BClearing, dehydration, then paraffin infiltration
   3. CParaffin infiltration, dehydration, then clearing
   4. DDehydration, paraffin infiltration, then clearing
   Show answer and explanation▾

   Answer: A. Dehydration, clearing, then paraffin infiltration

   Tissue is dehydrated in graded alcohols, cleared with a solvent such as xylene that is miscible with both alcohol and wax, and then infiltrated with paraffin. The other orders cannot work because wax does not mix with alcohol or water.

   Reference: Suvarna SK, Layton C, Bancroft JD, Bancroft's Theory and Practice of Histological Techniques, 8th ed. (2019)

9. Question 9Routine and Special Stainingmedium

   A pathologist requests a stain for iron in a liver biopsy. Which stain should be used?

   1. ACongo red
   2. BAlcian blue at pH 2.5
   3. CPerls Prussian blue
   4. DVerhoeff-van Gieson
   Show answer and explanation▾

   Answer: C. Perls Prussian blue

   Perls reaction turns ferric iron (hemosiderin) blue. Congo red is for amyloid, Alcian blue for acid mucins and Verhoeff-van Gieson for elastic fibers.

   Reference: Kiernan JA, Histological and Histochemical Methods: Theory and Practice, 5th ed. (2015)

10. Question 10Tissue Processing and Embeddinghard

    Which problem results from leaving tissue too long in absolute alcohol and xylene on an overnight processor?

    1. ASoft, swollen tissue that did not shrink and still contains water in the cells
    2. BDissolved nuclei that will not take up any hematoxylin during later staining
    3. CHard, brittle tissue that shrinks and cuts poorly
    4. DComplete loss of fixation with autolysis spreading from the center of the block
    Show answer and explanation▾

    Answer: C. Hard, brittle tissue that shrinks and cuts poorly

    Prolonged dehydration and clearing over-harden tissue, which becomes brittle and shatters on cutting. Soft tissue suggests under-processing, and nuclei and fixation are not lost by this step.

    Reference: Carson FL, Hladik C, Histotechnology: A Self-Instructional Text, 5th ed. (2021)

11. Question 11Routine and Special Stainingmedium

    Which special stain best highlights fungal organisms in tissue, with fungi appearing black on a green background?

    1. AGiemsa, in which fungi stain pink against a pale blue background
    2. BGrocott methenamine silver, which impregnates the fungal cell walls
    3. CVon Kossa, in which fungi appear brown against a clear yellow background
    4. DFontana-Masson, in which fungi stain deep red against a lilac background
    Show answer and explanation▾

    Answer: B. Grocott methenamine silver, which impregnates the fungal cell walls

    GMS reduces silver at fungal cell walls to give black organisms. Giemsa highlights H. pylori and blood cells, Von Kossa shows calcium and Fontana-Masson shows melanin.

    Reference: Kiernan JA, Histological and Histochemical Methods: Theory and Practice, 5th ed. (2015)

12. Question 12Routine and Special Stainingmedium

    A technologist wants to show collagen in blue or green and muscle in red. Which stain is correct?

    1. APAS with diastase
    2. BMasson trichrome
    3. COil Red O
    4. DAlcian blue
    Show answer and explanation▾

    Answer: B. Masson trichrome

    Masson trichrome stains collagen blue or green and muscle and cytoplasm red. PAS shows carbohydrates, Oil Red O shows lipid on frozen sections and Alcian blue shows mucin.

    Reference: Carson FL, Hladik C, Histotechnology: A Self-Instructional Text, 5th ed. (2021)

13. Question 13Specimen Reception, Fixation and Grossing Supportmedium

    A 20 mm thick piece of liver is placed in formalin and processed the next morning. What is the most likely problem?

    1. AOver-hardening of the surface only, while the center remains perfectly well fixed
    2. BExcess hematoxylin uptake that is caused by alkaline formalin in the container
    3. CPoor fixation of the center, because formalin penetrates only about 1 mm per hour
    4. DComplete loss of all cell nuclei through acid hydrolysis of the DNA overnight
    Show answer and explanation▾

    Answer: C. Poor fixation of the center, because formalin penetrates only about 1 mm per hour

    Formalin penetrates slowly (about 1 mm per hour), so thick tissue is unfixed centrally and then processed poorly. Surface hardening alone, hematoxylin excess and nuclear loss do not follow from this error.

    Reference: Suvarna SK, Layton C, Bancroft JD, Bancroft's Theory and Practice of Histological Techniques, 8th ed. (2019)

14. Question 14Specimen Reception, Fixation and Grossing Supportmedium

    For breast biopsies sent for ER, PR and HER2 testing, what are the ASCO/CAP fixation requirements?

    1. AFixation in ethanol for 2 hours with delay of up to 24 hours before fixation
    2. BFixation in Bouin fluid for 12 hours with no limit on delay
    3. CUnfixed transport on ice for 48 hours before processing
    4. DFixation in 10 percent neutral buffered formalin for 6 to 72 hours, with cold ischemia time of 1 hour or less
    Show answer and explanation▾

    Answer: D. Fixation in 10 percent neutral buffered formalin for 6 to 72 hours, with cold ischemia time of 1 hour or less

    ASCO/CAP guidance is 6 to 72 hours in neutral buffered formalin and cold ischemia of one hour or less, to preserve biomarkers. Ethanol, Bouin fluid and unfixed delay degrade antigens or nucleic acids.

    Reference: Allison KH et al., Estrogen and Progesterone Receptor Testing in Breast Cancer: ASCO/CAP Guideline Update, J Clin Oncol 2020;38:1346-1366

15. Question 15Specimen Reception, Fixation and Grossing Supportmedium

    A technologist receives a specimen container with a requisition showing a different patient surname from the container label. What should happen first?

    1. ARelabel the container using the requisition details, because the form is the legal record
    2. BProcess the specimen now to avoid delay and correct the record afterwards
    3. CDo not accession the specimen until the discrepancy is resolved with the sender
    4. DDiscard the specimen and automatically request a repeat biopsy from the clinic
    Show answer and explanation▾

    Answer: C. Do not accession the specimen until the discrepancy is resolved with the sender

    Mismatched identification must be resolved before accessioning, and the specimen must not be relabeled by the lab on assumption. Processing first risks wrong-patient reports, and discarding irreplaceable tissue is not the first step.

    Reference: Carson FL, Hladik C, Histotechnology: A Self-Instructional Text, 5th ed. (2021)

Last reviewed September 2026. Questions are original and written to the published exam blueprint; they are not taken from any real paper.