Staining Flashcards
why do we stain?
- overcome a lack of contrast in tissues
- identify cell types and structures
- identify specific molecules within a tissue
basic reaction of stains= attraction of _______
opposites
tissues that stain with a basic stain are?
basophilic
tissues that stain with an acidic stain?
acidophilic
what are the 6 basic dyes?
- methylene blue (blue)
- toludine blue ( blue)
- hematoxylin (blue)
- methyl green (green)
- basic fuschin
- pyronine G (red)
what are the 5 acidic dyes?
- eosin
- orange G
- aniline blue
- fast green
- acid fuschin
acidic dyes react with the _____ in solution rather than your section
alkali
basic dyes react with the _____ in solution rather than in your section
acid
how does hematoxylin react?
reacts like abasicdye with a purplish blue colour. It stains acidic, or basophilic, structure including the cell nucleus (which contains DNA and nucleoprotein), and organelles that contain RNA such as ribosomes and the rough endoplasmic reticulum
how does eosin stain?
acidicdye that is typically reddish or pink. It stains basic, or acidophilic, structures which includes the cytoplasm, cytoplasmic filaments, intracellular membranous components, cell walls and extracellular fibres
how does hematoxylin and eosin stain?
most common stain
- nuclei (blue basophilic) hematoxylin
- cytoplasm (pink acidophilic) eosin
what is the periodic acid Schiff reaction?
PAS- Most carbohydrates react with periodic acid to produce aldehydes, which convert the colourless Schiff reagent (Basic Fuchsin) to pink, or magenta.
trichromes are good for staining what?
connective tissue
how is connective tissue stained?
both employ a nuclear and cytoplasmic and a third stain for ecm
mallory’s trichrome staining can reveal ______
collagen
toludine blue is a _______ stain
metachromatic stain
toludine blue stains mast cells______ in a blue backround
purple
what is pararosaniline-toludine blue (PT)?
dye contains chromatin shades of purple and cytoplasm and collagn a lighter violet, penetrate plastic sections easier than H and E
_____________ -_______ is used for differential staining of cellular componants, particularly cytoplasmic granules
toludine blue
wright giemsa stain is for_______ _________
blood smears
describe the componants of the wright giemsa stain?
- azure blue stains white blood cell granules basophilic or neutrophilic
- eosin for RBC and eosinophilic WBC
- methylene blue stains nuclei
what is silver impregnation used for?
specificity provided by what silver is complexed and pH of staining solution, used to trace nerves, stain golgi and reticular fibres
lipids undergoe _______ because paraffin processing will extract lipids
cryosections
what are 4 lipid and fat stains?
- sudan black
- sudan IV (red )
- congo red
- Osmium tetroxide (black)
what is enzyme histochemistry?
Care during fixation to preserve enzyme activity. A specific enzyme catalyzed reaction is preformed with the section and the reaction product is visualized as an insoluble precipitated on the section.
what are some fluorescent stains?
acridine orange- binds nucleic acids
DAPI - binds DNA and fluorescein-phalloidin binds actin
what is microscopic autoradiography?
Radiolabeled molecule is injected into animal (e.g. 3H-fucose) and animal sacraficed.
Sections are coated with photographic emulsion, exposed and developed.
Microscopic autoradiography reveals location of newly synthesized glycoproteins containing fucose.
autoradiographs are?
are tissue preparations in which particles called silver grains indicate the cells or regions of cells in which specific macromolecules were synthesized just prior to fixation
what is in situ hybridization?
ISH ( localize the cells expressing a gene (DNA) or transcript (mRNA) of interest
_____ -______ are used to make specific antibodies by injecting with molecular of interest
host animals
antibodies are purified and used for either ____ or _______ immunolabeling of sections
direct or indirect
what are some benefits for confocal microscopy?
- reduced blurring of the image from light scatterign
- increased effective resolution
- improved signal to noise ratio
- clear examination of thick specimens
in TEM staining ______ fixation provides contrast
osmium
counter staining with heavy metals in done in TEM staining why?
provides electron density to section
- uranyl acetate
- lead citrate
stains are selective ( T or F)
TRUE
____ and _____ are important in clinical diagnostics
IHC and ISH