fresh tissue

54問 • 2年前

Kmyching

通報

問題一覧

used when a rapid diagnosis of a tissue is required.

FROZEN SECTION

without use of any chemical fixative

FREEZE DRYING

similar to freeze drying

FREEZE SUBSTITUTION

rapid freezing (- 160°C)

QUENCHING

removal of water in the form of ice (-40°C vacuum)

SUBLIMATION

Frozen tissue is submerged to________ (1% acetone) and dehydrated using absolute alcohol.

ROSSMAN’S FORMULA

o almost any microtome can be used o uses carbon dioxide

COLD KNIFE PROCEDURE

OPTIMUM WORKING TEMPERATURE = -18° to -20°C

CRYOSTAT PROCEDURE (COLD MICROTOME)

a refrigerated cabinet in which a modified microtome is housed.

CRYOSTAT

methods of freezing: best mounting media for cryostat

OPTIMAL CUTTING TEMPERATURE (OCT)

method of freezing:most rapid

LIQUID NITROGEN

methods of freezing: adequate for freezing small pieces of tissue EXCEPT muscle.

AEROSOL SPRAYS

Specimen size for processing

3X2 CM AND 3-5 MM THICK

FIRST AND MOST CRITICAL STEP Process of preserving cells and tissue constituents The tissue is preserved by preventing degeneration, putrefaction, decomposition and distortion

FIXATION/PRESERVATION

to preserve the morphologic and chemical integrity of the cell in as life-like manner as possible.

PRIMARY AIM

to harden and protect the tissue from the trauma of further handling.to harden and protect the tissue from the trauma of further handling.

SECONDARY AIM

Most important reaction for maintaining tissue morphology.

STABILIZATION OF PROTEINS

becomes part of the tissue by formation of cross links or complexes. Ex. Formalin, Hg, osmium tetroxide

ADDITIVE FIXATION

10% Formol saline, 10% Neutral buffered formalin (NBF), Heidenhain’s SuSa, Formol sublimate, Zenker’s solution, Zenker’s formol, Bouin’s solution, Brasil’s solution

MICROANATOMICAL FIXATIVE

ex. 10% Formol saline, Absolute Ethyl Alcohol, Newcomers fluid, Acetone (FANA)

HISTOCHEMICAL FIXATIVE

contains glacial acetic acid ex. Bouin’s fluid, Flemming’s fluid, Newcomers fluid, Carnoy’s fluid, Heidenhain’s SuSa (BFNCH). <4.6 pH

NUCLEAR FIXATIVE

ex. Helly’s fluid, Orth’s fluid, Regaud’s fluid, Flemming fluid without acetic acid, Formalin with post chroming (HORFF). >4.6 pH

CYTOPLASMIC FIXATIVE

fixatives containing mercuric chloride and potassium dichromate in cryostat section

Lipid Fixation

Alcoholic fixative for glycogen (Rossman’s fluid or cold absolute alcohol)

Carbohydrate Fixation

neutral buffered forma

Protein Fixation

gas produced by the oxidation of methyl alcohol. Concentrated solutions should not be neutralized (explosion) Stock solution: 37-40% Working solution: 10% (no buffer: unstable)

Formaldehyde (Formalin)

o White crystalline precipitates o Due to prolonged standing o Removed by: 10% METOH/filtration

Paraformaldehyde

Brown/black granular deposits that may obscure microscopic details

Acid formaldehyde hematin

diluted in 10% NaCl CNS

10% Formol Saline

Best general tissue fixative Best fixative for tissue containing iron granules w/ double phosphate buffer 1 mm/hr = rate of tissue penetration

10% Neutral Buffered Formalin (NBF) or PO4 buffered formalin

for routine post mortem tissues w/ HgCl2

Formol corrosive (formol sublimate)

Glutaraldehyde

EM: electron histochemistry & electron immunocytochemistry

Karnovsky’s paraformaldehyde – glutaraldehyde

Mixture w/ formaldehyde/formaldehyde

Acrolein

Lipids (frozen section)

Formol-calcium

HIGHLY EXPLOSIVE when dry. Excessive YELLOW STAINING of tissues. NEVER WASH IN WATER before dehydration. For glycogen (excellent)

PICRIC ACID FIXATIVE

recommended for fixation of embryos and pituitary biopsies

Bouin’s

Excellent fixative for glycogen and less messy then Bouin’s solution (excellent)

Brasil’s alcoholic picroformol fixative

It is normally used in conjunction with other fixatives to form a compound solution Solidifies at 17°C. Fixes & precipitates nucleoproteins, chromosomes, & chromatin material

GLACIAL ACETIC ACID

for fixing dry and wet smears (PBS and BM tissues)

Methyl alcohol

simple fixative incorporated with compound fixatives for better results, preserves but does not fix glycogen (Disadvantage: polarization)

Ethanol

for fixing touch preparation

Isopropyl alcohol

for fixing chromosomes, lymph glands and urgent biopsies (MOST RAPID;1-3 hours)

Carnoy’s fluid

to preserve sputum

Alcoholic Formalin (Gendre’s fixative)

for fixing mucopolysaccharides and nuclear proteins. Give better reaction in Fuelgen stain than Carnoys

Newcomer’s

Pale yellow powder which dissolves in water (up to 6% at 20°C) to form a strong oxidizing solution Inhibits hematoxylin

OSMIUM TETROXIDE

most common chrome-osmium acetic acid fixative used (FIXATIVE & DECALCIFYING AGENT), permanently fixes fat, for nuclear structures (excellent)

Flemmings solution

(improve cytoplasmic details) – recommended for mitochondria

Flemming’s solution without acetic acid

Precipitates proteins. Swelling effect counteract shrinkage by other fixatives. Weak decalcifying agent (softening effect)

TRICHLOROACETIC ACID

Used at ice cold temperature from -5°C to 4°C. For diffusible enzymes such as phosphatases and lipases. For fixing BRAIN TISSUE (Rabies Diagnosis)

ACETONE

Thermal coagulation of tissue proteins. For bacteriologic smears. Bacteriologic smears Microwave: 45-55°C

HEAT FIXATION

process of placing an already fixed tissue in a second fixative

SECONDARY FIXATION

fixation whereby a primarily fixed tissue is placed in Aqueos solution of 2.5-3% potassium dichromate

POST-CHROMATIZATION

Removing excess fixative

WASHING OUT

HISTOPATH (lesson 1)

Kmyching · 56問 · 2年前

HISTOPATH (lesson 1)

56問 • 2年前

Kmyching

bacte (microbial control)

Kmyching · 26問 · 2年前

bacte (microbial control)

Collection

Collection

Histopath (somatic death)

Kmyching · 25問 · 2年前

Histopath (somatic death)

Staphylococcus

Staphylococcus

lesson 1 (cell injury, adaptation and inflammation and repair)

Kmyching · 67問 · 2年前

lesson 1 (cell injury, adaptation and inflammation and repair)

biopsy

biopsy

autopsy

autopsy

decalcification

decalcification

DEHYDRATION AND CLEARING

Kmyching · 27問 · 2年前

DEHYDRATION AND CLEARING

27問 • 2年前

Kmyching

IMPREGNATION AND EMBEDDING

Kmyching · 21問 · 2年前

IMPREGNATION AND EMBEDDING

21問 • 2年前

Kmyching

TRIMMING AND SECTIONING

Kmyching · 35問 · 2年前

TRIMMING AND SECTIONING

35問 • 2年前

Kmyching

adhesive and staining

Kmyching · 43問 · 2年前

adhesive and staining

43問 • 2年前

Kmyching

COMMON STAINING SOLUTION

Kmyching · 19問 · 2年前

COMMON STAINING SOLUTION

OTHER STAINING

OTHER STAINING

H&E STAINING, MOUNTING, RINGING AND LABELLING

Kmyching · 19問 · 1年前

H&E STAINING, MOUNTING, RINGING AND LABELLING

IMMUNOHISTOCHEMISTRY

IMMUNOHISTOCHEMISTRY

EPITHELIAL TUMOR MARKERS

Kmyching · 5問 · 1年前

EPITHELIAL TUMOR MARKERS

5問 • 1年前

Kmyching

INTERMEDIATE FILAMENT MARKERS

Kmyching · 6問 · 1年前

INTERMEDIATE FILAMENT MARKERS

6問 • 1年前

Kmyching

NEUROENDOCRINE MARKERS and GERM CELL TUMOR MARKERS

Kmyching · 6問 · 1年前

NEUROENDOCRINE MARKERS and GERM CELL TUMOR MARKERS

6問 • 1年前

Kmyching

MESENCHYMAL TUMOR MARKERS

Kmyching · 5問 · 1年前

MESENCHYMAL TUMOR MARKERS

5問 • 1年前

Kmyching

問題一覧

used when a rapid diagnosis of a tissue is required.

FROZEN SECTION

without use of any chemical fixative

FREEZE DRYING

similar to freeze drying

FREEZE SUBSTITUTION

rapid freezing (- 160°C)

QUENCHING

removal of water in the form of ice (-40°C vacuum)

SUBLIMATION

Frozen tissue is submerged to________ (1% acetone) and dehydrated using absolute alcohol.

ROSSMAN’S FORMULA

o almost any microtome can be used o uses carbon dioxide

COLD KNIFE PROCEDURE

OPTIMUM WORKING TEMPERATURE = -18° to -20°C

CRYOSTAT PROCEDURE (COLD MICROTOME)

a refrigerated cabinet in which a modified microtome is housed.

CRYOSTAT

methods of freezing: best mounting media for cryostat

OPTIMAL CUTTING TEMPERATURE (OCT)

method of freezing:most rapid

LIQUID NITROGEN

methods of freezing: adequate for freezing small pieces of tissue EXCEPT muscle.

AEROSOL SPRAYS

Specimen size for processing

3X2 CM AND 3-5 MM THICK

FIRST AND MOST CRITICAL STEP Process of preserving cells and tissue constituents The tissue is preserved by preventing degeneration, putrefaction, decomposition and distortion

FIXATION/PRESERVATION

to preserve the morphologic and chemical integrity of the cell in as life-like manner as possible.

PRIMARY AIM

to harden and protect the tissue from the trauma of further handling.to harden and protect the tissue from the trauma of further handling.

SECONDARY AIM

Most important reaction for maintaining tissue morphology.

STABILIZATION OF PROTEINS

becomes part of the tissue by formation of cross links or complexes. Ex. Formalin, Hg, osmium tetroxide

ADDITIVE FIXATION

10% Formol saline, 10% Neutral buffered formalin (NBF), Heidenhain’s SuSa, Formol sublimate, Zenker’s solution, Zenker’s formol, Bouin’s solution, Brasil’s solution

MICROANATOMICAL FIXATIVE

ex. 10% Formol saline, Absolute Ethyl Alcohol, Newcomers fluid, Acetone (FANA)

HISTOCHEMICAL FIXATIVE

contains glacial acetic acid ex. Bouin’s fluid, Flemming’s fluid, Newcomers fluid, Carnoy’s fluid, Heidenhain’s SuSa (BFNCH). <4.6 pH

NUCLEAR FIXATIVE

ex. Helly’s fluid, Orth’s fluid, Regaud’s fluid, Flemming fluid without acetic acid, Formalin with post chroming (HORFF). >4.6 pH

CYTOPLASMIC FIXATIVE

fixatives containing mercuric chloride and potassium dichromate in cryostat section

Lipid Fixation

Alcoholic fixative for glycogen (Rossman’s fluid or cold absolute alcohol)

Carbohydrate Fixation

neutral buffered forma

Protein Fixation

gas produced by the oxidation of methyl alcohol. Concentrated solutions should not be neutralized (explosion) Stock solution: 37-40% Working solution: 10% (no buffer: unstable)

Formaldehyde (Formalin)

o White crystalline precipitates o Due to prolonged standing o Removed by: 10% METOH/filtration

Paraformaldehyde

Brown/black granular deposits that may obscure microscopic details

Acid formaldehyde hematin

diluted in 10% NaCl CNS

10% Formol Saline

Best general tissue fixative Best fixative for tissue containing iron granules w/ double phosphate buffer 1 mm/hr = rate of tissue penetration

10% Neutral Buffered Formalin (NBF) or PO4 buffered formalin

for routine post mortem tissues w/ HgCl2

Formol corrosive (formol sublimate)

Glutaraldehyde

EM: electron histochemistry & electron immunocytochemistry

Karnovsky’s paraformaldehyde – glutaraldehyde

Mixture w/ formaldehyde/formaldehyde

Acrolein

Lipids (frozen section)

Formol-calcium

HIGHLY EXPLOSIVE when dry. Excessive YELLOW STAINING of tissues. NEVER WASH IN WATER before dehydration. For glycogen (excellent)

PICRIC ACID FIXATIVE

recommended for fixation of embryos and pituitary biopsies

Bouin’s

Excellent fixative for glycogen and less messy then Bouin’s solution (excellent)

Brasil’s alcoholic picroformol fixative

It is normally used in conjunction with other fixatives to form a compound solution Solidifies at 17°C. Fixes & precipitates nucleoproteins, chromosomes, & chromatin material

GLACIAL ACETIC ACID

for fixing dry and wet smears (PBS and BM tissues)

Methyl alcohol

simple fixative incorporated with compound fixatives for better results, preserves but does not fix glycogen (Disadvantage: polarization)

Ethanol

for fixing touch preparation

Isopropyl alcohol

for fixing chromosomes, lymph glands and urgent biopsies (MOST RAPID;1-3 hours)

Carnoy’s fluid

to preserve sputum

Alcoholic Formalin (Gendre’s fixative)

for fixing mucopolysaccharides and nuclear proteins. Give better reaction in Fuelgen stain than Carnoys

Newcomer’s

Pale yellow powder which dissolves in water (up to 6% at 20°C) to form a strong oxidizing solution Inhibits hematoxylin

OSMIUM TETROXIDE

most common chrome-osmium acetic acid fixative used (FIXATIVE & DECALCIFYING AGENT), permanently fixes fat, for nuclear structures (excellent)

Flemmings solution

(improve cytoplasmic details) – recommended for mitochondria

Flemming’s solution without acetic acid

Precipitates proteins. Swelling effect counteract shrinkage by other fixatives. Weak decalcifying agent (softening effect)

TRICHLOROACETIC ACID

Used at ice cold temperature from -5°C to 4°C. For diffusible enzymes such as phosphatases and lipases. For fixing BRAIN TISSUE (Rabies Diagnosis)

ACETONE

Thermal coagulation of tissue proteins. For bacteriologic smears. Bacteriologic smears Microwave: 45-55°C

HEAT FIXATION

process of placing an already fixed tissue in a second fixative

SECONDARY FIXATION

fixation whereby a primarily fixed tissue is placed in Aqueos solution of 2.5-3% potassium dichromate

POST-CHROMATIZATION

Removing excess fixative

WASHING OUT