Sample type required: Deparaffinized and rehydrated tissue section (6–8 μm) on positively (+) charged slides
Preferred fixative: 10% neutral buffered formalin (NBF)
Control: Normal cerebral cortex
| Reagent | Time | Technical Notes |
| Phospohmolybdic acid-alcohol | 3 minutes | - Drain off excess fluid and place slides on a staining rack.
|
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.
|
| Crystal violet stain | 30 seconds | - Prepare fresh.
- Add stain while the sections are still covered in absolute alcohol chloroform.
|
| 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 air dry thoroughly.
|
| Differentiating solution* | 30 seconds | - Prepare fresh.
- Differentiate each slide INDIVIDUALLY.
|
| 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.
|
Post-staining procedure: The tissue section should be covered immediately.
Expected results:
Glial fibers - Purplish blue
Background - Pale blue to colorless
