Staining Procedures For Plastic Embedded Tissue
Verified at the Applications Laboratory of the Biomedical Division, Sorvall
Microtomes
SOLUTIONS:
| Acid Orcein Solution |
|
0.2g |
|
Orcein |
|
100.0ml |
|
Ethyl alcohol, 70% |
|
0.6ml |
|
Hydrochloric acid, concentrated
Solution is ready for staining immediately and improves on standing.
It is stable for many months. |
|
|
|
|
| Nocht's Azure-Eosin |
|
|
|
See procedure 2-4. |
STAINING PROCEDURE:
- Cut tissue at 2µm to 10µm. Thicker sections
give better three-dimensional picture.
- Heat slides on 60ºC to 80ºC hot plate for
10 minutes.
- Cool slides.
- Stain in acid orcein solution at 60ºC for
30-45 minutes.
- Rinse in running tap water for 15 minutes.
If sections begin to detach, blow dry, the refix on hot plate for 10 minutes.
Cool, then proceed. If sections are overstained, decolorize in absolute alcohol
or 0.1% hydrochloric acid alcohol, then tap water wash.
- Rinse in distilled water.
- Stain in Nocht's azure-eosin procedure (2-4) for 1 hour.
Blow dry.
Mount.
RESULTS:
| Nuclei |
|
deep blue |
| cytoplasm of epidermis, smooth muscle, and other cells |
|
light blue |
| collagen |
|
rose pink |
| elastic fibers |
|
dark brown |
| mast cell granules and many mucoid substances |
|
purple |
| eosinophilic granules and erythrocytes |
|
bright red |
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. |
