Staining Procedures For Plastic Embedded Tissue
Verified at the Applications Laboratory of the Biomedical Division,
Sorvall Microtomes
SOLUTIONS:
| Congo Red Stock Solution |
|
3.0g |
|
Congo red |
|
3.0g |
|
Sodium chloride |
|
1000.0ml |
|
Alcohol, 80% |
|
|
|
|
| Congo Red Working Solution |
|
50.0ml |
|
Congo red stock solution |
|
0.5ml |
|
Sodium hydroxide, 1%
Filter and use within 20 minutes. |
|
|
|
|
| Ammonia Water |
|
|
|
Add a few drops of ammonium hydroxide to tap water to raise
pH and insure proper staining. |
|
|
|
|
| Harris' Hematoxylin |
|
|
|
See formula in procedure
2-7, b. |
STAINING PROCEDURE:
- Section tissue at 6µm.
- Stain in working congo red solution for 10 minutes.
- Rinse in distilled water; blot dry.
- Stain in Harris' hematoxylin for 10 minutes.
- Rinse in tap water for 5 minutes.
- Dip in ammonia water until sections turn blue.
- Rinse in tap water for 5 minutes.
- Rinse in distilled water for 1 minute.
Blow dry.
Mount.
RESULTS:
| Amyloid |
|
red and shows dichromic metachromasia on polarization |
| nuclei |
|
blue |
Warning: Some of the chemicals
used for the staining procedures given in this section may be hazardous
if misused. For this reason, read and observe all warnings and cautions
provided by the manufacturer for each chemical before proceeding
with a staining procedure.
Note: In order to prevent sections from
loosening from the slides during staining, all sections should
be heat-fixed (60ºC to
100ºC) to the slides for a minimum of 2-5 minutes prior to staining,
preferably at the time the sections are mounted on the slides. |
