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HTL fixation

QuestionAnswer
Reactive site for formaldehyde and its effect on staining -NH2, (amino group). Leaves tissue basophilic, less receptive to acid dyes. e.g. reduced affinity for eosin
10% aqueous formalin constituents formaldehyde water
10% formal saline constituents formaldehyde 0.9% sodium chloride (adjust to isotonic) water
calcium formalin constituents formaldehyde calcium chloride (preserves phospholipids) water
Formalin ammonium bromide constituents Formaldehyde ammonium bromide (makes solution very acidic, pH~1.5) water
Acetate formalin constituents Formaldehyde sodium acetate (pH buffer to limit formation of formalin pigment) water
10% neutralized formalin constituents formaldehyde calcium/magnesium phosphate (antacid/neutralizing agents) water
10% neutral buffered formalin constituents Formaldehyde sodium phosphate mono and dibasic (pH buffers) water
Modified Millonig Formalin constituents Formaldehyde sodium phosphate monobasic sodium hydroxide water
Alcoholic formalin constituents Formaldehyde ethyl alcohol (dehydrates as it fixes, good for processor when time is short) water
Phosphate buffered paraformaldehyde constituents Paraformaldehyde sodium phosphate monobasic sodium hydroxide water
B5 constituents Mercuric chloride sodium acetate water (stock soln.) Formaldehyde (working soln.)
Bouin Solution constituents Picric acid acetic acid formaldehyde
Gendre solution constituents 95% ETOH saturated with picric acid Acetic acid formaldehyde
Hollande solution constituents Copper acetate (stabilize RBC, and granules of eosinophils and endocrine cells) picric acid formaldehyde acetic acid water
Zenker solution constituents Mercuric chloride potassium dichromate sodium sulfate water (stock solution) Acetic acid (working solution)
Helley Solution constituents Mercuric chloride potassium dichromate sodium sulfate water (stock) Formaldehyde (Working)
Orth Solution constituents Potassium dichromate sodium sulfate water formaldehyde
Zamboni (PAF) solution constituents Paraformaldehyde picric acid Sodium phosphate mono/dibasic water
Unbuffered Aqueous Zinc formalin constituents Zinc sulfate distilled water Formaldehyde
Alcoholic zinc chloride formalin constituents Zinc chloride water isopropyl alcohol formaldehyde
Carnoy solution constituents Absolute ethyl alcohol chloroform acetic acid
Clarke fluid constituents Absolute ethyl alcohol glacial acetic acid
Michael transport medium constituents Anhydrous citric acid ammonium sulfate N-ethylmaleimide Magnesium sulfate water
PBS stock buffer constituents sodium phosphate mono basic sodium phosphate dibasic sodium chloride
PBS-10% sucrose constituents Stock PBS sucrose water
Lugol iodine solution constituents Iodine potassium iodide distilled water
reactive/additive site for Osmium tetroxide Double bonds of unsaturated lipids and phospholipids
reactive/additive site for mercuric chloride Sulfhydryl groups Amino groups (if acidic)
mercuric chloride effect on staining/morphology enhances both acidic and basic stains; great nuclear detail because it is a nuclear coagulant, preserves organelles
Osmium tetroxide effect on staining and morphology poor staining with acidic dye, good staining with basic dyes excellent morphological preservation
reactive/additive sties of picric acid basic amino acids, causes basic the proteins to precipitate. this is the reason that acid dye staining is so effective following picric acid fixation: that basic proteins are well preserved, but acid proteins are not adequately fixed and may be removed
reactive/additive sites of potassium dichromate reactis with carboxyl and hydroxyl groups, increases the number of amino groups available for acid stains to bind
Reactive/additive site of zinc salts not listed in carson. Prevents crosslinking.
For good fixation, tissue should be no larger than: 2cm square, and no more than 4mm thick
Flemming Solution Constituents 2% osmium tetroxide 1% chromic acid glacial acetic acid
one indication of good use for zenker fluid is fuelgen reaction
Zenker fluid is not recommended for silver stains
Zenker fixation effects of acid/base staining decreased basophilia of nuclei, increaced acidophilia of cytoplasm
One disadvantage of osmium tetroxide interferes with many subsequent staining techniques
is potassium dichromate a suitable fixative for histochemistry? NO. it's affinity for carboxyl and hydroxyl groups renders it unsuitable.
The order of steps taken for gluteraldehyde fixation of EM specimens is: 1: Gluteraldehyde (~2hrs), 2: phosphate buffer wash, 3:sucrose/gum wash (is storage is required, skipped if processed immediately), 4: osmium postfixation
gendre's solution is recommended for what tissue component Glycogen
Created by: joshuajohnson7
 

 



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