Sample type required: Deparaffinized and rehydrated tissue section (6-8 μ) on positively (+) charged slides
Preferred fixative: 10% neutral buffered formalin (NBF)
Control: Normal cerebral cortex
Phospohmolybdic acid-alcohol | 3 minutes | - Drain off excess fluid and place slides on a staining rack.
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Absolute alcohol - chloroform mixture
| N/A | - Prepare fresh.
- Cover each section with this mixture while still on the staining rack.
- The tissue should become translucent.
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Crystal violet stain | 30 seconds | - Prepare fresh.
- Add stain while the sections are still covered in absolute alcohol - chloroform.
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10% potassium bromide | 1 minute | - Drain slides of absolute alcohol-chloroform mixture and crystal violet stain.
- Wash in potassium bromide.
- Blot each section and allow them to thoroughly air dry.
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Differentiating solution* | 30 seconds | - Prepare fresh.
- Differentiate each slide INDIVIDUALLY.
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Xylene | 3 changes | - Wash.
- Check macroscopically to ensure that the background is pale or colorless.
- Repeat the differentiating step(*) as needed to obtain this result.
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Post staining procedure: Tissue section should be coverlipped immediately.
Expected results:
Glial fibers - Purplish blue
Background - Pale blue to colorless