| Toluidine
Blue Staining Procedure
Literature
Purpose:
This standard operating procedure (SOP) establishes the standard
for staining mast cell granules using the Toluidine Blue staining
procedure.
Reagents
and Materials
Reagents
1% Toluidine Blue
Commercially
prepared by Lab Chem Inc. (Catalog# lc26165-1).
Expiration date provided by manufacturer.
Store at room temperature.
Materials
Tissue slides and control slides for special stain requested.
Microwave to bake slides.
Staining rack, 20 slide capacity.
Coverslips and synthetic mounting media.
Hazards
and Precautions:
See Material Safety Data Sheets.
Sample Preparations:
Paraffin tissue section slides cut at 4 microns.
Calibration
and Standardization:
A tissue section slide containing mast cells is stained with each
procedure.
Procedure:
1.
Deparaffinize and hydrate to distilled water.
2. 1% Toluidine Blue - 4 minutes.
3. Distilled water rinse.
4. Dehydrate, clear and coverslip.
Results:
Mast cell granules of most species - brilliant red to purple
Acid mucopolysaccharides - red to pink
Nuclei - shades of blue
Erythrocytes - green to yellow
Cherie
Chapman
James R. Turk
7/1/04
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