Histopathologic Techniques and Cytology Vocabulary

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Vocabulary flashcards covering Histopathologic Techniques and Cytology.

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108 Terms

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Teasing or Dissociation

A histopathologic technique where a tissue specimen is immersed in isotonic salt solution, dissected, and examined under a microscope.

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Squash Preparation or Crushing

A histopathologic technique where small tissue pieces are compressed between two slides and forcibly compressed with another slide or with a cover glass.

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Streaking

A histopathologic technique using an applicator stick or platinum loop to apply material in a direct or zigzag line on a slide.

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Spreading

A histopathologic technique where a portion of material is transferred to a slide and gently spread into a moderately thick film.

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Pull-Apart

A histopathologic technique utilized for preparing smears of thick secretions such as serous fluids, enzymatic lavage samples from the gastrointestinal tract and blood smears

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Touch Preparation or Impression Smear

A histopathologic technique where the surface of a freshly cut tissue is brought into contact and pressed onto a clean slide.

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Nitrogen

The most rapid freezing agent.

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Carbon Dioxide

A freezing agent used in a frozen microtome.

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Prions

Infectious agents that cause spongiform encephalopathies such as Creutzfeld Jacob disease (CJD), scrapie and madcow.

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Fixation

The first and most critical step in histotechnology.

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Primary Aim of Fixation

To preserve the morphological and chemical integrity of the cell in a life-like manner.

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Secondary Aim of Fixation

To harden and protect the tissue from the trauma of further handling, so that it is easier to cut during gross examination.

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Recommended Ratio of Fixative to Tissue

20 times the volume of tissue (10-20 times).

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Hollow Organs Fixation

Packed with cotton soaked in fixative and completely opened before immersion in fixing solution.

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Air-Filled Lungs Fixation

Lungs are covered with gauze to maintain it underwater.

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Eyes Fixation

Should not be dissected before they are fixed since this may lead to tissue collapse and wrinkling due to escape of vitreous humor.

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Brain Fixation

Suspended WHOLE in 10% buffered formalin for to 2 to 3 weeks to ensure fixation and hardening.

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Hard Tissues Fixation

Washed in running water overnight and immersed in 4% aqueous solution for 1 to 3 days (Lendrum’s technique).

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Concentrated solution of formaldehyde

Precipitates violent explosions (Should never be neutralized).

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Removal of Brown or Black Crystalline Formalin Deposits

Alcoholic picric acid or 1% KOH in 80% alcohol

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Removal of White Paraformaldehyde Deposits

Filtration or by addition of 10% methanol

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Methanol

Formaldehyde preservative that prevents decomposition to formic acid or paraformaldehyde precipitation.

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Microanatomic Fixatives

10% formol saline and 10% neutral buffered formalin.

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Cytoplasmic Fixatives

Flemming’s fluid without acetic acid and Helly’s.

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Nuclear Fixatives

Flemming’s, Carnoy’s, Bouin’s, Newcomer’s, and Heidenhain’s Susa.

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Formaldehyde Waste Disposal

Can be recycled through distillation, drain disposal after detoxification, or disposed of by a licensed hauler.

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Lipid Fixation

Formaldehyde or Aldehyde fixatives.

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Formaldehyde

Recommended for NERVOUS TISSUE PRESERVATION

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Alcoholic Formalin (Gendre’s) Fixative

Fixes sputum since it coagulates mucus.

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Mercuric Chloride

Most common metallic fixative Fixative of choice for tissue photography

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Dezenkerization

Alcoholic iodine Removal of excessive mercuric fixative Iodine then sodium thiosulfate then water

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Zenker’s Fluid

Recommended for fixing small pieces of liver, spleen, connective tissue fibers and nucleus

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Zenker-Formol

Excellent microanatomic fixative for pituitary gland, bone marrow and blood containing organs

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B5 Fixative

Recommended for bone marrow biospsy

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Orth’s Fluid

Recommended for degenerative processes and tissue necrosis Demonstrates Rickettsiae and other bacteria

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Regaud’s Fluid

Recommended for demonstration of chromatin, mitochondria, mitotic figures, Golgi body and RBCs

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Heidenhain’s Susa

Recommended for tumor biopsy of the skin

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Glacial Acetic Acid

Fixative that imparts swelling of tissues

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Alcohol Fixatives

Fixatives that produce cell lysis and is ideal for small tissue fragments

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Methyl Alcohol

Excellent for fixing dry and wet smears, blood smears and BM smears

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Isopropyl Alcohol

Used for touch preparation fixation and for Wright Giemsa staining procedures

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Carnoy’s Fluid

Recommended for fixing chromosomes, lymph glands and urgent biopsy Used for fixing brain tissues for diagnosis of rabies

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Newcomer’s Fluid

Recommended for fixing mucopolysaccharides and nuclear proteins; nuclear and histochemical fixative

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Flemming’s Solution

With ACETIC ACID: Most common chrome osmium acetic acid fixative; recommended for nuclear fixation Without ACETIC ACID: Made up only of chromic and osmic acid; recommended for structures particularly mitochondria

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Acetone

Recommended for study of water diffusible enzymes especially for phosphatases Used at ice cold temperature -5 to 4 C Used in fixing brains for rabies diagnosis

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Post Chromatization

Secondary fixation with 2.5 to 3% potassium dichromate and acts as a mordant for better staining effects and to aid in cytologic preservation of tissues

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Advantage of Microwave Fixation

Tissue is heated right through the block in a very short time; thereby allowing the study of cellular processes that proceed very rapidly.

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Enzyme Histochemistry Fixatives

4% Formaldehyde or formol saline

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Fixatives for Electron Microscopy

  1. OSMIUM TETROXIDE 2. Glutaraldehyde 3. Paraformaldehyde 4. Karnovsky’s paraformaldehyde-glutaraldehyde mixture
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Decalcification

Complete removal of calcium salt from the tissues like bone and teeth and other calcified tissues following fixation

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Measuring Extent of Decalcification

  1. Physical or Mechanical Test 2. X-Ray or Radiological Test 3. Chemical Method 4. Bubble Method
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Ethanol

Recommended for routine dehydration of tissues; best dehydrating agent

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Methyl Alcohol

Toxic dehydrating agent; primarily employed for blood and tissue films

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Butyl Alcohol

Utilized in plant and animal microtechniques; slow dehydrating agent

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Xylene

Most commonly used clearing agent

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Paraffin Wax

Simplest, most common and best embedding medium for routine tissue processing

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Substitutes for Paraffin Wax

  1. Paraplast 2. Embeddol 3. Bioloid 4. Tissue mat 5. Ester wax 6. Water soluble waxes (carbowax)
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Paraplast

Highly purified paraffin and synthetic plastic polymers, mp of 56 to 57 C

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Embeddol

Synthetic wax substitute similar to paraplast with a mp of 56-58 C; less brittle and less compressible than paraplast

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Bioloid

Semisynthetic wax recommended for embedding eyes

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Tissue Mat

Product of paraffin containing rubber, same property as paraplast

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Ester Wax

Lower mp (46-48 C) but is harder than paraffin

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Carbowax

Polyethylene glycol; miscible and soluble in water, hence does not require dehydration and clearing of tissues

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Celloidin

Purified form of nitrocellulose soluble in many solvents

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Thin (2%), Medium (4%) or Thick (8%)

Solution of cellulose dissolved in equal parts of ether and alcohol

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Gelatin Impregnation

Water soluble, does not require dehydration and cleaning Rarely used except when tissues are subjected to histochemical and enzyme preparation

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Orientation

Process by which tissue is arranged in precise position in the mold during embedding on the microtome before cutting and on the slide before staining

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Rocking Microtome

Simplest microtome used in cutting serial sections of large blocks of paraffin embedded tissues by Paldwell Trefall

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Rotary Microtome

Most common type Used for cutting paraffin embedded sections Rotary microtome may be a. manual (completely manipulated by the operator) b. semi-automated (one motor to advance either the fine or coarse HAND WHEEL) c. fully-automated (two motors that drive both the fine and the coarse advance HAND WHEEL)

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Freezing Microtome

Releases CO2 that freezes the tissues Used for cutting unembedded frozen tissues

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Biconcave Knife

Recommended knife for cutting paraffin embedded sections on a rotary microtome

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Plane Wedge Knife

Recommended knife for frozen sections for cutting extremely large and tough specimens embedded in paraffin blocks, using a base sledge type or sliding microtome

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Diamond Glass Knives

Knives for electron microscopy

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Mayer’s Egg Albumin

most common adhesive for surgical sections

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Rapid Diagnosis

Primary application of frozen section

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Cold Knife Procedure

One method of preparing frozen sections

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Cryostat Procedure

One method of preparing frozen sections

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Cryostat

Also known as a cold microtome

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Direct Staining

Process of giving color to the sections by using simple aqueous or alcoholic dye solutions

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Indirect Staining

Process whereby action of the dye is intensified by adding another agent (eg. MORDANT)

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Mordant

Serves as a link between the tissue and the dye

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Regressive Staining (Differentiation)

Tissue is first overstained and the excess is removed or decolorized

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Orthochromatic Staining

Color shades are similar to the color of the dye itself

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Metachromatic Staining

Differentiate particular substances by staining them with a color that is different from that of the stain

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Counterstaining

Application of a different color or stain to provide contrast and background

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Best Vital Dye

Neutral Red

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Most common method of staining for microanatomical studies of tissues

Hematoxylin and Eosin

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Chromophore

coloring property, capable of producing visible colors of synthetic dyes

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Auxochrome

Dyeing property; retains color of synthetic dyes

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Hematoxylin and Eosin

Most common method of staining for microanatomical studies of tissuesResults: NUCLEUS: blue to blue black CYTOPLASM: pink

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Mounting

To avoid distortion of the image, the refractive index of the mountant should be as near as possible to that of the glass which is 1.518

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ETHER ALCOHOL

Best fixative for cytology

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Sputum Collection

At least three (3) consecutive MORNING specimens

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Ascites, Peritoneal fluids and Cell Suspensions

Optimum amount is 20 to 30 ml effusion Cells remain viable for up to 4 days if REFRIGERATED (4°C), do not freeze.

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Papanicolaou (Pap' s)

Consists of three (3) stains: 1 Hematoxylin: stains the nucleus 2. Orange green (OG6): stains the cytoplasm of mature cells (superficial cells) 3. Eosin azure (EA 36/50/65): stains the cytoplasm of immature cells (parabasal and intemediate cells)

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Direct Technique (Immunohistochemistry)

Conjugate the primary antibody directly to the label such as fluorochrome or horseradish peroxidase

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Indirect Technique (Immunohistochemistry)

Two (2) or three (3) step procedure that involves application of unconjugated primary antibody, followed by a labeled antibody directed against the first antibody

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Chromogen for Peroxidases (Immunohistochemistry)

AMINOETHYLCARBAZOL or DIAMINOBENZIDINE

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Squamous Epithelium

Stratified; Lining oral cavity, Epiglottis, Esophagus, Anus, Cervix, Vagina, Vulva, glans penis, Cornea

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Sarcoma

Malignant tumor of CONNECTIVE TISSUE (mesenchymal) origin