Histology Fundamentals

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Questions and Answers

Match the following steps in histology with their descriptions:

Fixation = Preserve the tissue and prevent structural change. Embedding = Supporting medium for tissue Sectioning = Cutting the tissue into thin slices Staining = Adding color to the tissue for examination

Match the following terms with their definitions:

Formalin = A mixture of formaldehyde gas and water Formaldehyde = A gas used in fixation Fixation = Preserving tissue from decay Autolysis = Breakdown of tissue by its own enzymes

Match the following materials with their uses:

10% N.B.Formalin = Most common fixative in histology Methyl alcohol = Added to formaldehyde to make formalin Neutral buffer = Added to formalin to make it suitable for fixation Water = Used to dilute 100% formalin

Match the following steps with their purposes:

<p>Embedding = To support the tissue for sectioning Sectioning = To cut the tissue into thin slices Staining = To add color to the tissue for examination Fixation = To preserve the tissue from decay</p> Signup and view all the answers

Match the following statements with their corresponding reasons:

<p>Formalin is cheap = It is widely used in histology Formalin penetrates tissue rapidly = It preserves the natural state of tissue 10% Formalin is used = It is the best fixative to preserve the natural state 90 ml of water is added = To dilute 10 ml of 100% Formalin</p> Signup and view all the answers

Match the following terms with their descriptions:

<p>Histology = Study of tissue Pathologist = A doctor who examines tissue slides Histology department = Prepares slides of tissue for examination Autolysis = Breakdown of tissue by its own enzymes</p> Signup and view all the answers

Match the following steps with their results:

<p>Fixation = Tissue is preserved from decay Embedding = Tissue is supported for sectioning Sectioning = Tissue is cut into thin slices Staining = Tissue is colored for examination</p> Signup and view all the answers

Match the following materials with their concentrations:

<p>100% Formalin = 37% solution of formaldehyde gas and water 10% Formalin = 10 ml of 100% Formalin plus 90 ml of water 37% solution = Formaldehyde gas solution Water = Diluent for 100% Formalin</p> Signup and view all the answers

Match the following steps with their description in the histology process:

<p>Step 3 = Wax infiltration Embedding = Adding an external base to the tissue Microtomy = Cutting sections of the tissue Labelling Slides = Identifying the slides with a label</p> Signup and view all the answers

Match the following chemicals with their usage in the histology process:

<p>Xylene = Wax solvent Formalin = Fixative Ethyol alcohol = Dehydrating agent Hematoxylin = Stain for tissue</p> Signup and view all the answers

Match the following equipment with their usage in the histology process:

<p>Microtome = Cutting sections of the tissue Cryostat = Preparing frozen sections HistoCore PELORIS 3 = Automated tissue processing Hot water bath = Flattening sections</p> Signup and view all the answers

Match the following stains with their usage in the histology process:

<p>Hematoxylin and Eosin = Common stain Oil Red O = Stain for lipids Orcein = Stain for elastic in arteries or cartilage Toluidine blue = Stain for H pylori</p> Signup and view all the answers

Match the following specimen preparation methods with their primary uses:

<p>Cytocentrifugation Preparation = Transfering biological cells onto a microscope slide ThinPrep Preparation = Cervical smear preparation Cell Block Preparation = Capturing remaining specimen in the centrifuge tube Histology Preparation = Not mentioned</p> Signup and view all the answers

Match the following steps with their purpose in the histology process:

<p>Dehydration = Removing water from the tissue Clearing = Removing dehydrating agent Wax infiltration = Replacing the dehydrating agent with wax Embedding = Adding an external base to the tissue</p> Signup and view all the answers

Match the following steps with their order in the histology process:

<p>Wax infiltration = Step 3 Embedding = Step 4 Microtomy = Step 5 Labelling Slides = Step 6</p> Signup and view all the answers

Match the following tasks with the role of an MLA in Histology/Cytology:

<p>Prepares specimens for histology = MLA task Performs routine tissue processor maintenance = MLA task Analyzes patient samples = Not an MLA task Files and retrieves stained slides = MLA task</p> Signup and view all the answers

Match the following equipment with their uses:

<p>Cytocentrifuge = Transferring sedimentable particles onto a microscope slide ThinPrep 2000 Processor = Automated slide preparation for cervical cancer testing Microscope = Analyzing prepared slides Centrifuge = Separating particles of different densities</p> Signup and view all the answers

Match the following disposal methods with their respective chemicals:

<p>Down the drain = Ethyol alcohol Fractional distillation = Xylene Neutralizing agents = Formalin Not recyclable = Paraffin wax</p> Signup and view all the answers

Match the following materials with their uses:

<p>Histogel = Hardening cell blocks Cytology fixed spray = Fixing microscope slides Formalin = Preserving tissue samples Pap test = Cervical cancer testing</p> Signup and view all the answers

Match the following steps with their purpose in the histology process:

<p>Staining = Adding color to the tissue Permanent mounting = Adding a coverslip to the slide Labelling Slides = Identifying the slides with a label Hydration = Rehydrating the tissue</p> Signup and view all the answers

Match the following equipment with their benefits in the histology process:

<p>HistoCore PELORIS 3 = Automated tissue processing Cryostat = Rapid preparation of frozen sections Microtome = Cutting sections of the tissue Hot water bath = Flattening sections</p> Signup and view all the answers

Match the following steps with their procedures:

<p>Insert broom into larger opening of the vial = Specimen collection Place the cap on the vial = Securing the specimen Send to lab in a specimen bag = Transporting the specimen Mounting slides = Preparing slides for analysis</p> Signup and view all the answers

Match the following steps with their outputs in the histology process:

<p>Microtomy = Ribbon of sections Staining = Colored tissue Labelling Slides = Labeled slides Permanent mounting = Finished slide</p> Signup and view all the answers

Match the following tasks with their corresponding procedures:

<p>Stains and mounts pap slides = Cytology procedure Prepares specimens for histology = Histology procedure Performs routine staining procedures = Histology procedure Analyzes patient samples = Not mentioned</p> Signup and view all the answers

Match the following materials with their corresponding procedures:

<p>Tissue paper = Cell block preparation Cassette = Storing and transporting specimen blocks Formalin = Preserving tissue samples Microscope slides = Analyzing prepared slides</p> Signup and view all the answers

Match the following fixatives with their characteristics:

<p>10% formalin = Fixes 4 mm piece of tissue in 8 hrs Bouin's solution = Contains picric acid Glutaraldehyde = Used for electron microscopy Alcohols = Good for cytology</p> Signup and view all the answers

Match the following factors with their effects on fixation:

<p>Temperature = Increases the rate of diffusion of the fixative Time = The longer the better in most cases Penetration rate = Depends on the fixative Specimen dimensions = 1-4 mm thickness</p> Signup and view all the answers

Match the following tasks with their corresponding responsibilities:

<p>Prepares specimens for cytology = MLA task Files and retrieves stained slides = MLA task Analyzes patient samples = Not an MLA task Assists in the cutting room = MLA task</p> Signup and view all the answers

Match the following steps with their descriptions in tissue processing:

<p>Dehydration = Removal of H2O using a ascending series of alcohols Clearing = Displacing ethanol with an intermediate solvent Wax infiltration = Infiltrating the tissue with paraffin wax Embedding = Not mentioned in the passage</p> Signup and view all the answers

Match the following materials with their uses in tissue processing:

<p>Alcohols = Dehydration Xylene = Clearing Paraffin wax = Wax infiltration Ethanols = Not mentioned in the passage</p> Signup and view all the answers

Match the following types of tissues with their characteristics:

<p>Routine specimens = Examples include appendix Small tissue biopsy = Examples include colon bx Urgent specimens = Examples include breast bx Cytology blocks = Different color</p> Signup and view all the answers

Match the following labelling methods with their descriptions:

<p>Manual method = Causes problems such as penmanship issues Cassette Labeler = A machine that etches numbers on the cassette Barcode = Not mentioned in the passage Handwriting = Not mentioned in the passage</p> Signup and view all the answers

Match the following components with their places in the labelled cassette:

<p>Year = First part of the label Surgical specimen number = Second part of the label Lab number = Third part of the label Specimen number = Fourth part of the label</p> Signup and view all the answers

Match the following gross examination products with their descriptions:

<p>Document = Written record of the examiner's findings Tissue blocks = Portions of tissue sealed in plastic cassettes Gross description = Not mentioned in the passage Microscopic examination = Not mentioned in the passage</p> Signup and view all the answers

Match the following fixatives with their components:

<p>Bouin's solution = Picric acid Zenker's fixative = Mercuric chloride Glutaraldehyde = Not mentioned in the passage Sodium dihydrogen phosphate = Not mentioned in the passage</p> Signup and view all the answers

Match the following steps with their order in tissue processing:

<p>Dehydration = First step Clearing = Second step Wax infiltration = Third step Embedding = Not mentioned in the passage</p> Signup and view all the answers

Match the following types of cytology specimens with their descriptions:

<p>Gynaecological = Cervical cancer demonstrates distinct pre-cancerous cellular changes that may be detected by the pap smear Non-gynaecological = Specimens obtained from the respiratory, urinary or gastro-intestinal systems Fine Needle Aspirations = Specimens obtained from any site using a fine needle Cytology = The study of cells in terms of structure, function and chemistry</p> Signup and view all the answers

Match the following stains with their uses:

<p>Papanicolaou stain = For nuclear details in gynecologic cytology Romanowsky-type stain = For air-dried gynecologic preparations Hematoxylin = For staining cell nuclei blue Eosin Y = For staining cytoplasm pink</p> Signup and view all the answers

Match the following cytology procedures with their descriptions:

<p>Papanicolaou staining = Involves five dyes in three solutions Romanowsky staining = Used to differentiate cells for microscopic examination Cytology = The study of cells in terms of structure, function and chemistry Pap smear = The most common test done in cytology</p> Signup and view all the answers

Match the following specimen collection methods with their descriptions:

<p>LBC Collection Vial = A method of collecting specimens for cytology Rover Cervex Brush = A device used to collect patient samples Cooper Spatula/Brush Combo = A combination of tools used to collect patient samples Glass slides = No longer used for collecting smears</p> Signup and view all the answers

Match the following cytology specimen types with their examples:

<p>Cervical/Vaginal Smears = Gynaecological specimen Sputum = Non-gynaecological specimen Body Fluids = Non-gynaecological specimen Fine Needle Aspirations = Breast, lung, salivary glands, thyroid, liver, pancreas and lymph nodes</p> Signup and view all the answers

Match the following cytology procedures with their steps:

<p>Papanicolaou staining = Involves five dyes in three solutions Specimen collection = Remove the top 2D barcode from the vial and place on the bottom right corner of the Cytology requisition Romanowsky staining = Used to differentiate cells for microscopic examination Cytology = The study of cells in terms of structure, function and chemistry</p> Signup and view all the answers

Match the following cytology specimen types with their collection methods:

<p>Gynaecological = Using a broom and rotating 5 times in a clockwise direction Non-gynaecological = Using a Cooper Spatula/Brush Combo Fine Needle Aspirations = Using a fine needle Cytology = The study of cells in terms of structure, function and chemistry</p> Signup and view all the answers

Match the following cytology procedures with their purposes:

<p>Pap smear = To look for cancerous changes to cells Cytology = The study of cells in terms of structure, function and chemistry Papanicolaou staining = To stain cell nuclei blue Romanowsky staining = To differentiate cells for microscopic examination</p> Signup and view all the answers

Match the following cytology specimen types with their locations:

<p>Cervical/Vaginal Smears = Cervical canal Sputum = Respiratory system Body Fluids = Pericardial, pleural, peritoneal and joint fluids Fine Needle Aspirations = Breast, lung, salivary glands, thyroid, liver, pancreas and lymph nodes</p> Signup and view all the answers

Match the following cytology procedures with their requirements:

<p>Papanicolaou staining = Five dyes in three solutions Specimen collection = Patient label with First Name, Last Name and PHIN Romanowsky staining = Air-dried gynecologic preparations Cytology = The study of cells in terms of structure, function and chemistry</p> Signup and view all the answers

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Study Notes

Histology

  • The main function of a histology department is to prepare slides of tissue from various sources for pathologists to examine.
  • Preparation of histological sections involves:
    • Preservation of tissue through fixation to prevent structural changes.
    • Embedding the tissue in a supporting medium.
    • Sectioning the tissue.
    • Staining the tissue.

Fixation

  • Fixation is a critical step in histological section preparation to preserve tissue from decay.
  • The most common fixative is 10% neutral buffered formalin.
  • 10% neutral buffered formalin is cheap and penetrates tissue rapidly to preserve the natural state.
  • Formalin is made by mixing 40% formaldehyde with 100ml of water.

Factors Affecting Fixation

  • Temperature: Increasing the temperature of fixation increases the rate of diffusion of the fixative into the tissue and speeds up the rate of chemical reaction.
  • Time: The longer the fixation, the better in most cases.
  • Penetration rate: Depends on the fixative.
  • Specimen dimensions: Optimal thickness is 1-4 mm.

Alternative Fixatives

  • Bouin's solution: Contains picric acid.
  • Zenker's fixative: Contains mercuric chloride.
  • Glutaraldehyde: Used for electron microscopy.
  • Alcohols: Good for cytology.

Gross Description

  • The initial step in examining a clinical specimen.
  • Produces two end products: a written document and a set of tissue blocks.
  • The success of the final histological diagnosis depends on the skill of the professional performing the gross description.

Tissue Processing

  • Dehydration: Removes water from the tissue using a series of alcohols (70%, 95%, 100%).
  • Clearing: Uses an intermediate solvent to remove ethanol from the tissue.
  • Wax infiltration: Infiltrates the tissue with a suitable histological wax.

Embedding

  • Places the tissue in a mold with liquid embedding material, which is then hardened.
  • Creates a stable block that can be cut into thin sections.

Microtomy

  • Cuts sections of the tissue imbedded in the wax block using a microtome.
  • Produces a series of ribbons at a thickness of 3-5 um.

Labelling Slides

  • Labels slides with the same information as the block:
    • Year
    • S (if surgical specimen)
    • Lab number
    • Specimen number
    • Sub number

Staining

  • Most common stain is Hematoxylin and Eosin (H&E).
  • Removes wax from the sections using a wax solvent.
  • Hydrates the sections using descending alcohols and water.
  • Stains the sections with H&E.

Permanent Mounting

  • Dehydrates the slides with ascending alcohols and xylene.
  • Covers the slides with a mountant and a glass coverslip.
  • Removes excess mounting fluid using xylene.

Frozen Sections

  • A rapid alternative to embedding in paraffin wax.
  • Uses a cryostat to prepare and stain the tissue.
  • Tissue is unfixed.

Cytology

  • Definition: The study of cells in terms of structure, function, and chemistry.
  • Objectives:
    • Define cytology.
    • List the three types of cytology specimens.
    • Describe the different types of procedures used in cytology.
    • Describe the most common stain used in cytology.

Types of Cytology Specimens

  • Gynaecological: Cervical cancer demonstrates distinct pre-cancerous cellular changes.
  • Non-gynaecological: Obtained from respiratory, urinary, or gastrointestinal systems.
  • Fine needle aspirations: Obtained from various sites using a fine needle.

Stains Used in Cytology

  • Papanicolaou stain: Used for gynecologic cytology and fixed non-gynecological samples.
  • Romanowsky-type stain: Used for air-dried gynecologic preparations.

Specimen Preparation

  • Methods:
    • Cytocentrifugation preparation (Cytospin).
    • ThinPrep preparation: Mainly used for cervical smears.
    • Cell block preparation: Captures remaining specimen in the centrifuge tube.

Role of a Medical Laboratory Assistant (MLA) in Histology/Cytology

  • Prepares specimens for histology and cytology.
  • Labels tissue cassettes and slides.
  • Performs routine tissue processor maintenance and reagent changes.
  • Files and retrieves stained slides and paraffin blocks.
  • Assists in the cutting room.

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