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HTL fixation
| Question | Answer |
|---|---|
| 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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