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Histology ASCP
ASCP Histology Prep: STAINING
Question | Answer |
---|---|
If overstain w/ methylene blue counterstain in acid-fast procedure, you remove the excess color by ____ | differentiating in 95% alcohol or in acid-alcohol and restain |
stains that detect fungi | PAS for fungi; Grocott's methanamine silver nitrate; Gridley |
what color is a positive result for H. pylori using the Giemsa stain? | dark blue |
color of GRAM NEGATIVE organism using the Gram stain? | RED (neg-red) |
what are the names of the acid fast stains | Kinyoun/ Fite/ Ziehl Neelsen/ Auramine-Rhodamine |
H. pylori detection stains are | Diff Quik/Giemsa/Alcian Yellow-Toluidine Blue/Steiner |
What is the genus name for the bacteria identified with acid-fast stain. | Mycobacteria |
What color is the background stained with acid fast stains? | blue |
What color does spirochetes stain? | BLACK |
What step must be added to the fite acid fast stain? | xylene peanut oil solution to deparaffinize in order to protect leprosy capsule |
What stains the leprosy organism? | Fite acid-fast stain |
What are the steps of the acid fast stain procedure? | Carbol fuchsin - differetiate w/acid alcohol- wash in running water- counterstain in methylene blue- rinse with water, dehydrate, clear, coverslip |
What color does H.Pylori stain using Diff Quik? | blue violet |
what are the steps of the gram stain? | crystal violet- gram's iodine - safranin or basic fuchsin |
What color do GRAM POSITIVE organisms stain using Gram stain? | blue or violet |
What color signifies a positive result for acid fast? | red |
What does Gram stain differentiate between? | gram(+) and gram(-) organisms |
what color does H. pylori stain with Steiner? | black |
What stain detects spirochetes? | Steiner/Wathin-Starry |
With GMS (Grocott's methanimine silver nitrate), what color does fungi stain? | BLACK |
What is the most recognized pigment? | melanin |
most common exogenous pigments | carbon/iron/lipochromes |
What pigment can bind silver? | melanin |
3 most common artifactual pigments | formalin/mercury/chromium |
5 common endogenous pigments | melanin/hemosiderin/urates/bile/lipofuchsin |
Van Gieson stain results | blue/black nuclei; RED COLLAGEN |
PAS is used for ________ mucosubstances | NEUTRAL |
In metachromatic staining, ___ can be varied to differ acid mucins | pH |
To suppress background staining in Congo Red procedure, _________________ is used. | Sodium Chloride |
Alcian Blue/ PAS is used to differentiate between ________. | Acid & Neutral Mucosubstances |
is birerefringet | formalin |
PAS diastese | shows sites in the tissue where the glycogen was removed (digested). |
Colloidal Iron binds with acid mucosubstances in what reaction and stains blue? | Prussian blue reaction. |
FITE stains what? | Mycobacterium (RED) |
Prepare a 10% solution of 37%-40% formalin with | 9 parts water adn one part stock solution formalin. |
Paraformaldehyde is used in ____ labs to yield pure solution of formaldehyde, which do not contain _________. | EM labs/Methanol |
______ helps to prevent polymerization | methanol |
cross links quickly but binds slowly | formaldehyde |
Black acid hematin forms when formalin pH is below ____ in blood rich tissues. | 6.0 |
greatest binding action of formaldehyde occurs at ____ to ___ pH | 7.5 to 8.0 pH |
Formaldehyde does NOT fix carbohydrates but traps them ______... | inside the fixed and stabilized proteins. |
Contains Sodium Chloride/DI/Formaldehyde | Formaldehyde Saline: isotonic but may produce formalin pigment |
Contains Calcium Chloride/DI/Formaldehyde | Calcium Formaldehyde: for PHOSPHOLIPIDS |
Contains Sodium Acetate/DI/Formaldehyde | Acetate Formaldehyde: for Phospholipids but cases PSEUDOCALCIFICATION |
Contains Calcium or Magnesium Carbonate/DI/Formaldehyde | 10% Neutralized Formalin: used widely but becomes acidic almost immediately. |
Contains Sodium Phosphate monobasic & dibasic/DI/Formaldehyde | 10% Neutral Buffered Formalin: for routine formalin fixation. ph is 6.8 |
Contains Sodium Phosphate Monobasic/Sodium Phosphate HYDROXIDE/DI/Formaldehyde | Modified Millonig: used in EM on long stored tissue |
contains Absolute Ethyl Alcohol/DI/Formaldehyde | Alchoholc Formalin: store tissue indefinitely |
Formalin PEL = 0.75ppm in __ hours | 8 hours |
Formalin STEL = 2 ppm in ___ | 15 minutes |
Contains Sodium Phosphate monobasic/25% glutaraldehyde/DI | Glutaraldehyde (pH = 7.2-7.3) |
Coagulant, produces shrinkage, additive, black pigment NOT preventable | Mercuric Chloride (not used with frozens, inhibits freezing/corrosive/avoid use with metals) |
EM use as post-fixative after aldehyde fixation. Cell membranes become electron dense. use very thin sections. | Osmium tetroxide |
____ | Gridley fungus stain |
TWA = 0.002ppm | Osmium tetroxide |
coagulant of necleoproteins, leaves DNA soluble, acts as stain and fixative, decalcifies, softens but shrinks too | Picric Acid (avoid for DNA and RNA) |
non-coagulant, great for MITOCHONDRIA identification procedures | Potassium Dichromate |
replacement for mercury, preserves antigenicity, may replace NBF, makes formalin pigment when unbuffered, superior nuclear detail | Zinc Salts |
fixative of hematopoitic and lymphoreticular tissues for beautiful nuclear detail (over NBF) | B5 |
mercuric chloride/formaldehyde/sodium acetate (formalin pigment) | B5 |
picric acid/glacial acetic acid/formaldehyde | Bouin |
red blood cells are lysed, formalin pigment may form, good for trichrome stain, remove yellow color with 50-70% alc, excellent for biopsy tissues, not for EM | Bouin |
95% alcohol w/picric acid/formaldehyde/glacial acetic acid | GENDRE |
alcoholic Boiun derivative which is great for carbohydrates (GLYCOGEN)remove excess picric with 80% alcohol | GENDRE |
copper acetate/picric acid/acetic acid/formaldehyde | HOLLANDE |
decalcifies small specs of bone, good for gastric biopsies, cupric acetate stabilizes RBC and eosinophils and endocrine cells. | HOLLANDE |
Any fixative containing formalin can yield _______. | formalin pigment, when the ph is lowered or if solution is unbuffered. |