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Vocabulary flashcards covering Histopathologic Techniques and Cytology.
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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.
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.
Streaking
A histopathologic technique using an applicator stick or platinum loop to apply material in a direct or zigzag line on a slide.
Spreading
A histopathologic technique where a portion of material is transferred to a slide and gently spread into a moderately thick film.
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
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.
Nitrogen
The most rapid freezing agent.
Carbon Dioxide
A freezing agent used in a frozen microtome.
Prions
Infectious agents that cause spongiform encephalopathies such as Creutzfeld Jacob disease (CJD), scrapie and madcow.
Fixation
The first and most critical step in histotechnology.
Primary Aim of Fixation
To preserve the morphological and chemical integrity of the cell in a life-like manner.
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.
Recommended Ratio of Fixative to Tissue
20 times the volume of tissue (10-20 times).
Hollow Organs Fixation
Packed with cotton soaked in fixative and completely opened before immersion in fixing solution.
Air-Filled Lungs Fixation
Lungs are covered with gauze to maintain it underwater.
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.
Brain Fixation
Suspended WHOLE in 10% buffered formalin for to 2 to 3 weeks to ensure fixation and hardening.
Hard Tissues Fixation
Washed in running water overnight and immersed in 4% aqueous solution for 1 to 3 days (Lendrum’s technique).
Concentrated solution of formaldehyde
Precipitates violent explosions (Should never be neutralized).
Removal of Brown or Black Crystalline Formalin Deposits
Alcoholic picric acid or 1% KOH in 80% alcohol
Removal of White Paraformaldehyde Deposits
Filtration or by addition of 10% methanol
Methanol
Formaldehyde preservative that prevents decomposition to formic acid or paraformaldehyde precipitation.
Microanatomic Fixatives
10% formol saline and 10% neutral buffered formalin.
Cytoplasmic Fixatives
Flemming’s fluid without acetic acid and Helly’s.
Nuclear Fixatives
Flemming’s, Carnoy’s, Bouin’s, Newcomer’s, and Heidenhain’s Susa.
Formaldehyde Waste Disposal
Can be recycled through distillation, drain disposal after detoxification, or disposed of by a licensed hauler.
Lipid Fixation
Formaldehyde or Aldehyde fixatives.
Formaldehyde
Recommended for NERVOUS TISSUE PRESERVATION
Alcoholic Formalin (Gendre’s) Fixative
Fixes sputum since it coagulates mucus.
Mercuric Chloride
Most common metallic fixative Fixative of choice for tissue photography
Dezenkerization
Alcoholic iodine Removal of excessive mercuric fixative Iodine then sodium thiosulfate then water
Zenker’s Fluid
Recommended for fixing small pieces of liver, spleen, connective tissue fibers and nucleus
Zenker-Formol
Excellent microanatomic fixative for pituitary gland, bone marrow and blood containing organs
B5 Fixative
Recommended for bone marrow biospsy
Orth’s Fluid
Recommended for degenerative processes and tissue necrosis Demonstrates Rickettsiae and other bacteria
Regaud’s Fluid
Recommended for demonstration of chromatin, mitochondria, mitotic figures, Golgi body and RBCs
Heidenhain’s Susa
Recommended for tumor biopsy of the skin
Glacial Acetic Acid
Fixative that imparts swelling of tissues
Alcohol Fixatives
Fixatives that produce cell lysis and is ideal for small tissue fragments
Methyl Alcohol
Excellent for fixing dry and wet smears, blood smears and BM smears
Isopropyl Alcohol
Used for touch preparation fixation and for Wright Giemsa staining procedures
Carnoy’s Fluid
Recommended for fixing chromosomes, lymph glands and urgent biopsy Used for fixing brain tissues for diagnosis of rabies
Newcomer’s Fluid
Recommended for fixing mucopolysaccharides and nuclear proteins; nuclear and histochemical fixative
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
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
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
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.
Enzyme Histochemistry Fixatives
4% Formaldehyde or formol saline
Fixatives for Electron Microscopy
Decalcification
Complete removal of calcium salt from the tissues like bone and teeth and other calcified tissues following fixation
Measuring Extent of Decalcification
Ethanol
Recommended for routine dehydration of tissues; best dehydrating agent
Methyl Alcohol
Toxic dehydrating agent; primarily employed for blood and tissue films
Butyl Alcohol
Utilized in plant and animal microtechniques; slow dehydrating agent
Xylene
Most commonly used clearing agent
Paraffin Wax
Simplest, most common and best embedding medium for routine tissue processing
Substitutes for Paraffin Wax
Paraplast
Highly purified paraffin and synthetic plastic polymers, mp of 56 to 57 C
Embeddol
Synthetic wax substitute similar to paraplast with a mp of 56-58 C; less brittle and less compressible than paraplast
Bioloid
Semisynthetic wax recommended for embedding eyes
Tissue Mat
Product of paraffin containing rubber, same property as paraplast
Ester Wax
Lower mp (46-48 C) but is harder than paraffin
Carbowax
Polyethylene glycol; miscible and soluble in water, hence does not require dehydration and clearing of tissues
Celloidin
Purified form of nitrocellulose soluble in many solvents
Thin (2%), Medium (4%) or Thick (8%)
Solution of cellulose dissolved in equal parts of ether and alcohol
Gelatin Impregnation
Water soluble, does not require dehydration and cleaning Rarely used except when tissues are subjected to histochemical and enzyme preparation
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
Rocking Microtome
Simplest microtome used in cutting serial sections of large blocks of paraffin embedded tissues by Paldwell Trefall
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)
Freezing Microtome
Releases CO2 that freezes the tissues Used for cutting unembedded frozen tissues
Biconcave Knife
Recommended knife for cutting paraffin embedded sections on a rotary microtome
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
Diamond Glass Knives
Knives for electron microscopy
Mayer’s Egg Albumin
most common adhesive for surgical sections
Rapid Diagnosis
Primary application of frozen section
Cold Knife Procedure
One method of preparing frozen sections
Cryostat Procedure
One method of preparing frozen sections
Cryostat
Also known as a cold microtome
Direct Staining
Process of giving color to the sections by using simple aqueous or alcoholic dye solutions
Indirect Staining
Process whereby action of the dye is intensified by adding another agent (eg. MORDANT)
Mordant
Serves as a link between the tissue and the dye
Regressive Staining (Differentiation)
Tissue is first overstained and the excess is removed or decolorized
Orthochromatic Staining
Color shades are similar to the color of the dye itself
Metachromatic Staining
Differentiate particular substances by staining them with a color that is different from that of the stain
Counterstaining
Application of a different color or stain to provide contrast and background
Best Vital Dye
Neutral Red
Most common method of staining for microanatomical studies of tissues
Hematoxylin and Eosin
Chromophore
coloring property, capable of producing visible colors of synthetic dyes
Auxochrome
Dyeing property; retains color of synthetic dyes
Hematoxylin and Eosin
Most common method of staining for microanatomical studies of tissuesResults: NUCLEUS: blue to blue black CYTOPLASM: pink
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
ETHER ALCOHOL
Best fixative for cytology
Sputum Collection
At least three (3) consecutive MORNING specimens
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.
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)
Direct Technique (Immunohistochemistry)
Conjugate the primary antibody directly to the label such as fluorochrome or horseradish peroxidase
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
Chromogen for Peroxidases (Immunohistochemistry)
AMINOETHYLCARBAZOL or DIAMINOBENZIDINE
Squamous Epithelium
Stratified; Lining oral cavity, Epiglottis, Esophagus, Anus, Cervix, Vagina, Vulva, glans penis, Cornea
Sarcoma
Malignant tumor of CONNECTIVE TISSUE (mesenchymal) origin