Special stains
What is important to remember about Oil red O?
- Cut tissue sections at 8-10microns - Do not take the sections through xylene or alcohol - coverslip with aqueous mounting media
What are alternative lipid stains?
- Osmonium tetroxide - Sudan Black
What is used as the decolorizer in ZN?
Acid alcohol
What is Zielh Neelsen used to stain and what is the mechanism of staining?
Acid fast bacilli (TB), ionic bonding
What is congo red used to stain and what is its mechanism of staining?
Amyloid, hydrogen bonding
What is Halls used for and what is the mechanism of staining?
Bile stain, histochemical reaction
If Van geison is used as a counter stain, how would tissue components stain?
Cytoplasm, muscle and RBC's stains yellow from the picric acid Collagen stains red from the acid fuchsin
What is Alcian blue/PAS used for?
Differentiates between neutral and acid mucins
What is perls Prussian blue used for and what is the mechanism of staining?
Ferric iron, histochemical stain
What is the histochemical reaction in Halls bile?
Fouchets reagent= ferric chloride and TCA
In a GMS stain, if the chromic acid step is forgotten, what will be the result?
Fungi will not stain if present (false negative)
What stains are argentaffin silver/metal impregnation?
GMS Masson fontana
What would you use as a control for Halls stain?
Gallbladder, liver
What stains are argyrophilic silver/metal impregnation?
Gordon and Sweets retic, Warthin Starry and Von Kossa
In a gram twort what is stained by the twort solution?
Gram negative organisms and nuclei stain red from the neutral red and cytoplasm and RBCs stain green from the fast green
If the Iodine step in a Gram Twort stain is missed, what will the result be?
Gram positive organisms will not retain the crystal violet and will be stained with the counterstain. Gram positive- False negative Gram negative- false positive
In Kinyoun what is used to melt the waxy capsule of the bacteria and allow staining?
Higher concentrations of reagents
What is important about the Wade fites staining process?
Instead of dewaxing and hydrating, the slides go through a mixture of peanut oil and xylene because leprosy cell wall is fragile
What is the Toluidine Blue stain used to demonstrate?
Mast cells
What is Masson Fontana used to stain and what is the mechanism of staining?
Melanin stain, argentaffin silver impregnation
What is Oil red O used for and what is the mechanism of staining?
Neutral lipids, selective solubility
What is Tworts working solution made of?
Neutral red and fast green
Does GMS have a reducing reagent?
No, it is argentaffin
Does Masson Fontana have a reducing reagent?
No, it is argentaffin (the tissue reduces)
If tissue is suspected to be positive for asbestos, what would be a good stain to use?
Perls Prussian blue
What is Van Geison made of?
Picric acid and acid fuchsin
What is the Schmorals solution made out of?
Potassium ferricyanyde and ferric chloride
What are the reagents in order used in Masson Fontana?
Potassium permanganate Ammoniacle silver solution Gold chloride Sodium thiosulfate Nuclear fast red
What reagent is the bleacher in Masson Fontana? What does this mean?
Potassium permanganate or hydrogen peroxide, breaks the phenolic ring structure of melanin and removes the melanin pigment
What is Alcian blue used for and what is the mechanism of staining?
Stains acid mucins, ionic bonding
What is VonKossa used for and what is the mechanism of staining?
Stains calcium, argyrophilic silver impregnation
What is Warthin Starry used to stain and what is the mechanism of staining?
Stains spirochetes and H.pylori, argyrophilic silver impregnation
What is important about sections to be stained with congo red?
Tissue must be cut at 8-10 microns
What stains use turnbulls reaction?
Turnbulls blue (ferrous iron) and Schmorals (melanin)
Does Warthin Starry have a reducing reagent?
Yes, hydroquinone.
What is Wade Fites used to stain and what is the mechanism of staining?
acid fast bacilli of leprosy, ionic bonding
What is carbol fuchsin made of?
basic fuchsin, phenol and alcohol
What would be a control used for VonKossa?
bone
What would be a good control for PPB?
bone marrow, spleen
At pH of 2.5 what does alcian blue demonstrate?
both carboxylated and sulfated acid mucins
What must be done to the gms slides if chromic acid is used as the oxidizer?
chromic acid has to be removed with sodium metabisulfate or sodium bisulfate
What is used as the oxidizer in gms?
chromic acid or periodic acid
What is a good control slide for alcian blue?
colon, small intestine, appendix
What would be good controls for mucicarmide?
colon, small intestine, appendix
What is Gomori Aldehyde Fuchsin stain used for?
elastin fibers, mast cell granules, acid mucins
In Zielh Neelsen what is used to melt the waxy capsule of the bacteria and allow staining?
heat
What is the mechanism of staining for the gram twort?
ionic bonding
In alcian blue/PAS what does Schiffs reagent stain?
neutral mucins
What are the reagents that make up the Turnbull's blue solution?
potassium ferricyanide and HCl
What is perls working solution made of?
potassium ferrocyanayde and HCl
What colour do the acid fast bacilli stain in ZN?
red, stained by the carbol fuchsin
What would be a good control for Masson Fontana?
skin
What is a good control for Schmorals?
skin, melanin, tissue with argentaffin granules
If you do not have diastase to do a PASD stain, what can you do instead?
spit on the slide
In alcian blue/PAS what does alcian blue reagent stain?
stains acid mucins
What is Mucicarmide used for and what is the mechanism of staining?
stains epithelial acid mucins (corboxylated and sulfated but not neutral), empirical stain
What does Turnbulls blue stain and what is the mechanism of staining?
stains ferrous iron, histochemical stain
What is GMS used to stain and what is the mechanism of staining?
stains fungi, argentaffin silver impregnation
What is Schmorals used for and what is the mechanism of staining?
stains melnin, reducing substances, argentaffin granules and lipofuscin, histochemical stain
At pH of 1.0 what does alcian blue demonstrate?
sulfated mucosubstances
What is Alizarin red S used for?
used to stain small amounts of calcium