Calling all pathology professionals, lab scientists and students! Ready to put your expertise to the test? Our free histopathology quiz is designed to challenge and expand your mastery of histopathology techniques, from staining protocols to slide interpretation. In this engaging histopathology practice test, you'll tackle realistic lab histopathology questions that reveal your strengths and highlight areas for improvement. Whether you're prepping for certification or just curious about advanced pathology techniques quiz, you'll gain practical insights and confidence with each question. Ready to dive in? Click to start our histopathology quiz or explore a broader pathology techniques quiz now - your lab bench awaits your verdict!
What is the primary purpose of hematoxylin and eosin (H&E) staining in histopathology?
To embed tissues in paraffin wax
To visualize tissue morphology and cellular detail
To stain lipids selectively
To decalcify bone samples
Hematoxylin stains nuclei blue-purple and eosin stains cytoplasm pink, providing excellent contrast for tissue architecture and cellular details. It is the standard stain in diagnostic histopathology laboratories worldwide. This combination allows pathologists to assess cell morphology and identify pathological changes. More on H&E staining.
Which fixative is most commonly used for routine histopathology specimens?
10% neutral buffered formalin
Glutaraldehyde
Bouin's solution
Absolute ethanol
10% neutral buffered formalin (NBF) is the most widely used fixative because it crosslinks proteins, preserving tissue architecture and preventing autolysis. The buffering prevents acid formation that can damage nucleic acids. It offers good morphological preservation compatible with many downstream techniques. Details on formalin fixation.
What is the main purpose of tissue processing before embedding?
To cut thin sections for slides
To fix tissues with crosslinking agents
To decalcify mineralized tissues
To dehydrate and impregnate tissues with paraffin wax
Tissue processing removes water through graded alcohols and replaces it with a hydrophobic medium like paraffin wax. This step is essential to allow sectioning of thin, uniform slices. Proper processing ensures good morphology and stain uptake. Learn about tissue processing.
What is a microtome used for in histopathology?
Embedding tissue in paraffin
Scanning slides electronically
Staining tissue sections
Cutting thin sections of embedded tissue
A microtome holds paraffin blocks and slices them into thin sections (usually 3 - 5 µm thick) for mounting on microscope slides. Precision sectioning is critical for clear microscopic interpretation. Different microtomes (rotary, sliding, cryostat) are used depending on the sample. More about microtomes.
What is the typical thickness of paraffin-embedded tissue sections for routine light microscopy?
4 - 5 micrometers
1 - 2 micrometers
10 - 12 micrometers
20 - 25 micrometers
Sections of 4 - 5 µm provide optimal resolution of cellular details under the light microscope while maintaining tissue integrity. Thicker sections can obscure morphology, and thinner sections may tear or fold. Standardizing thickness is vital for reproducible staining and diagnosis. Section thickness guidance.
Why is deparaffinization necessary before staining paraffin-embedded tissue sections?
To remove paraffin so stains can penetrate the tissue
To fix the tissue further
To decalcify embedded bone
To digest proteins for antigen retrieval
Paraffin is hydrophobic and must be removed with xylene or substitutes so aqueous staining solutions can access tissue components. Proper deparaffinization ensures uniform staining and avoids patchy results. It is followed by rehydration through graded alcohols to water. Deparaffinization protocols.
Which chemical is most commonly used for deparaffinization in histology labs?
Xylene
Methanol
Hydrochloric acid
Acetic acid
Xylene is a nonpolar solvent that dissolves paraffin efficiently, facilitating its removal from tissue sections. Although toxic, it remains the standard in many labs or is replaced by less hazardous xylene substitutes. Xylene use requires fume hoods and proper disposal. Xylene in histology.
What does the term 'grossing' refer to in histopathology?
Embedding tissues in wax
Staining tissues with hematoxylin and eosin
Macroscopic examination and dissection of specimens
Microscopic examination of sections
Grossing is the initial step where a pathologist or histotechnologist inspects the specimen macroscopically, describes features, and dissects representative sections. Proper grossing ensures diagnostic areas are sampled. It is documented in a gross description for the pathology report. Grossing techniques.
What is the main purpose of antigen retrieval in immunohistochemistry?
To enhance fluorescence microscopy
To deparaffinize the section
To expose epitopes masked by formalin crosslinks
To decalcify mineralized tissue
Formalin fixation creates protein crosslinks that can mask antigenic sites. Antigen retrieval breaks these crosslinks (by heat or enzymes) to restore epitope accessibility for antibody binding. Proper retrieval significantly improves immunostaining sensitivity. Principles of antigen retrieval.
How does formalin primarily fix tissue?
By dissolving cellular membranes
By forming methylene bridges between proteins
By precipitating proteins with alcohol
By oxidizing lipids
Formaldehyde reacts with amino acid side chains to form methylene bridges (-CH2-), crosslinking proteins and stabilizing cellular structures. These covalent bonds preserve morphology and prevent autolysis. Excess fixation time can reduce antigenicity. Mechanism of formalin fixation.
Which histochemical stain is most specific for detecting glycogen in tissue sections?
Giemsa
Periodic acid - Schiff (PAS)
Alcian blue
Masson's trichrome
PAS oxidizes vicinal diols in glycogen to aldehydes, which react with Schiff reagent to yield a magenta color. It is the gold standard for visualizing glycogen and other carbohydrate-rich structures. Control diastase elimination confirms specificity. Details on PAS stain.
Which fixative is most appropriate for electron microscopy specimens?
10% neutral buffered formalin
Methanol
Bouin's solution
Glutaraldehyde
Glutaraldehyde provides stronger and more stable crosslinking than formalin, preserving ultrastructure for electron microscopy. It fixes both proteins and lipids effectively, maintaining membrane integrity. Specimens often undergo secondary fixation with osmium tetroxide. Glutaraldehyde in EM.
What is the purpose of decalcification in bone histology?
To fix the specimen
To enhance immunostaining
To remove calcium salts for sectioning
To stain osteoid matrix
Decalcification uses acids or chelating agents to dissolve calcium phosphate deposits in bone, making the tissue soft enough for microtome sectioning. Proper decalcification preserves cellular details while preventing blade damage. Over-decalcification can harm morphology and antigens. Guide to decalcification.
What is the function of a cryostat in histopathology?
Cutting fresh or frozen tissue sections at low temperatures
Dehydrating sections before staining
Performing immunohistochemistry
Embedding tissue in paraffin wax
A cryostat maintains a cold chamber (usually around - 20 °C) to keep tissue frozen for rapid sectioning without paraffin embedding. Frozen sections are used in intraoperative diagnoses and enzyme histochemistry. The speed of cryosectioning allows same-day results. About cryostats.
Which artifact is commonly associated with rapid freezing of tissue samples?
Paraffin embedding folds
Formalin pigment deposition
Ice crystal formation causing vacuoles
Martius scarlet bleeding
Ice crystals form during freezing if cooling is too slow, disrupting cellular structures and creating clear spaces (vacuoles). Proper snap-freezing techniques (e.g., isopentane in liquid nitrogen) minimize ice artifact. Artifacts can obscure diagnostic features. Freezing artifact guidance.
In immunohistochemistry for estrogen receptor (ER) detection, what is the role of the primary antibody?
Block non-specific binding
Decalcify the tissue
Provide the chromogenic reaction
Bind directly to the estrogen receptor antigen
The primary antibody specifically recognizes and binds to the estrogen receptor epitope on tissue sections. A labeled secondary antibody or detection system then binds the primary antibody to produce a visible signal. Specificity of the primary antibody is critical for accurate results. IHC antibody principles.
What is the main advantage of laser capture microdissection (LCM) in histopathology research?
Enhancing histochemical stain intensity
Isolating specific cells from tissue for molecular analysis
Automating slide scanning
Embedding tissues without paraffin
LCM allows precise excision of selected cells or regions directly from microscopic slides, preserving morphology while enabling downstream DNA, RNA, or protein studies. It minimizes contamination from surrounding tissues. This technique bridges histology with molecular diagnostics. More on LCM.
Which system is commonly used for signal amplification in immunohistochemistry?
Decalcification
Direct fluorescent tagging
Paraffin embedding
Biotin-streptavidin complex
The biotin - streptavidin (or avidin) method uses multiple biotin molecules on the secondary antibody, amplifying binding sites for streptavidin-linked enzymes. This enhances chromogenic or fluorescent signal sensitivity. It is widely adopted in pathology labs. Biotin-streptavidin amplification.
How can you distinguish glycogen from neutral mucin using histochemical stains?
Use H&E stain at higher pH
Use PAS with diastase digestion
Use silver impregnation
Use Masson's trichrome
PAS stains both glycogen and mucin, but pre-treatment with diastase digests glycogen, eliminating its PAS positivity while mucin remains stained. This differential method distinguishes the two substances in tissue sections. PAS-diastase technique.
Why is neutral buffered formalin preferred over unbuffered formalin?
It accelerates decalcification
It enhances paraffin infiltration
It improves antigen retrieval
It prevents formalin pigment and acid artifacts
Unbuffered formalin can become acidic over time, producing formalin pigment deposits and causing tissue acid hydrolysis. Neutral buffering maintains pH around 7, preserving tissue morphology and preventing artifacts. This stability is crucial for reliable staining. Buffered vs unbuffered formalin.
What is a key difference between immunofluorescence (IF) and immunoperoxidase (IHC) techniques?
IF uses fluorescent tags, IHC uses enzyme-mediated chromogens
IF requires paraffin embedding, IHC does not
IF visualizes DNA, IHC visualizes proteins
IF only works on frozen tissue, IHC only on paraffin
Immunofluorescence employs fluorophore-conjugated antibodies viewed under a fluorescence microscope, offering high sensitivity but requiring specialized equipment. Immunoperoxidase uses enzymes (like HRP) to catalyze a colorimetric reaction visible in bright-field microscopy. Each has its strengths in diagnostic practice. Comparing IF and IHC.
Which method employs microwave heating in a citrate buffer for antigen retrieval?
Protease-induced epitope retrieval (PIER)
Heat-induced epitope retrieval (HIER)
Microwave-assisted decalcification
Xylene-based retrieval
HIER uses microwave, pressure cooker, or water bath heating in buffers (commonly citrate pH 6) to reverse formalin crosslinks. It restores antigenicity for improved antibody binding. Proteolytic methods (PIER) use enzymes instead but have different specificity. HIER techniques.
What is the primary role of positive and negative controls in immunohistochemistry?
Enhance chromogen intensity
Reduce background autofluorescence
Validate staining specificity and technique reliability
Accelerate fixation
Positive controls confirm that the antibody and detection system work properly by staining a known antigen-positive tissue. Negative controls (omitting primary antibody) ensure that observed staining is specific and not due to nonspecific binding or reagent contamination. Controls are essential for accurate diagnostic interpretation. IHC control standards.
What is the typical resolution (pixel size) for diagnostic whole slide imaging scanners?
0.25 micrometers per pixel
10 micrometers per pixel
5 micrometers per pixel
50 micrometers per pixel
High-resolution digital pathology scanners capture images at ~0.25 µm/pixel to match or exceed the resolution of standard light microscopes. This detail allows pathologists to view subcellular structures and fine cellular morphology. Lower resolutions are insufficient for routine diagnostics. Whole slide imaging resolution.
What does double staining in immunohistochemistry allow you to do?
Measure antigen quantity by colorimetric ratio
Detect two antigens simultaneously with different chromogens
Stain the same antigen twice for intensity
Perform IHC and ISH on the same slide
Double staining uses two primary antibodies raised in different species and distinct chromogens, enabling visualization of two antigens in one tissue section. It provides spatial relationship data between markers, crucial in research and diagnostics. Careful antigen retrieval and control selection are required. Double IHC staining methods.
How does MALDI imaging mass spectrometry contribute to histopathology?
Decalcifies bone for sectioning
Maps spatial distribution of proteins and lipids in tissue sections
Replaces H&E staining with autofluorescence
Provides electron microscopic images
MALDI (Matrix-Assisted Laser Desorption Ionization) imaging MS ionizes biomolecules directly from tissue sections, generating spatial distribution maps of proteins, lipids, and metabolites. It offers molecular insights co-registered with histology, aiding biomarker discovery. The technique complements traditional staining. MALDI imaging overview.
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AI Study Notes
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Study Outcomes
Understand Tissue Staining Methods -
By completing this histopathology quiz, you will grasp the principles and applications of essential staining techniques such as H&E, PAS, and immunohistochemistry.
Analyze Microscopic Tissue Features -
You will learn to differentiate normal and abnormal cellular structures and apply histopathology techniques to interpret visual findings accurately.
Apply Laboratory Protocols Correctly -
This section covers best practices for sample preparation, slide handling, and safety procedures in response to common lab histopathology questions.
Evaluate Troubleshooting Strategies -
After the quiz, you will be able to identify and resolve common errors in staining, fixation, and sectioning to improve diagnostic quality.
Assess Your Diagnostic Proficiency -
End-of-quiz scoring and case-based questions will help you benchmark your skills against standard histopathology practice test scenarios.
Cheat Sheet
Tissue Fixation Fundamentals -
Proper fixation preserves cellular architecture by cross-linking proteins and preventing autolysis; formalin (10% neutral buffered formalin) is the gold standard as noted by the College of American Pathologists. Quick immersion and a fixative-to-tissue ratio of at least 10:1 ensure complete penetration. To remember fixation order, use the mnemonic "F.A.B.E.R." (Formalin, Alcohol, Buffer, EtOH, Rinse).
Essential Staining Protocols -
Hematoxylin and eosin (H&E) staining remains the cornerstone of histopathology techniques, with hematoxylin binding nucleic acids (blue-purple) and eosin highlighting cytoplasmic proteins (pink). According to the Journal of Histotechnology, optimal pH ranges for hematoxylin mordanting are 2.7 - 2.9. Special stains like PAS for glycogen or Masson's trichrome for connective tissue add diagnostic depth in your histopathology practice test.
Precision in Microtomy -
Consistent section thickness (typically 3 - 5 µm) is crucial for uniform staining intensity and is recommended by the International Academy of Pathology to avoid overstaining or poor resolution. Proper blade angle (5 - 10°) and regular blade replacement prevent chatter artifacts. Remember the phrase "Thin, clean, keen": thin sections, clean blade, keen eye for quality when tackling lab histopathology questions.
Artifact Recognition -
Artifacts such as tissue folds, knife marks, and air bubbles can mimic pathological changes and mislead diagnosis; PubMed studies emphasize training to distinguish real lesions from prep errors. Gentle floating of sections on a 40 - 45 °C water bath and proper mounting techniques reduce wrinkles. Use the mnemonic "F.A.B." (Folds, Air bubbles, Bookmarks) to recall common issues during your pathology techniques quiz.
Microscopic Interpretation Skills -
Develop systematic scanning of low to high power fields to assess architecture before cellular detail, a method endorsed by the World Health Organization tumor classification guidelines. Incorporate immunohistochemistry panels judiciously to confirm lineage markers and avoid false positives. For a confidence boost, practice with virtual slide libraries from academic research repositories to sharpen skills for your histopathology quiz.
