2-2. Gomori's Aldehyde Fuchsin (Refs 2,
5)
Staining Procedures For Plastic Embedded Tissue
Verified at the Applications Laboratory of the Biomedical Division, Sorvall
Microtomes
SOLUTIONS:
| Aldehyde Fuchsin Solution |
|
1.0g |
|
Basic fuchsin |
|
200.0ml |
|
Alcohol, 70% |
|
2.0ml |
|
Hydrochloric acid, concentrated |
|
2.0ml |
|
Paraldehyde (trimer acetaldehyde)
Let stand at room temperature for 1-2 days or until stain is deep purple
in color. Staining solution is good for about 4 weeks at room temperature
and for over 6 months in the refrigerator. Fresh solutions stain quicker
than used ones. |
|
|
|
|
| Lugol's Iodine |
|
1.0g |
|
Iodine |
|
2.0g |
|
Potassium iodide |
|
100.0ml |
|
Distilled water |
|
|
|
|
| 0.5% Sodium Bisulfite |
|
0.5g |
|
Sodium bisulfite |
|
100.0ml |
|
Distilled water |
|
|
|
|
| Heidenhain's Azan Method
or Van Gieson Method |
|
|
|
See procedure 2-3 or 2-12,
respectively. |
|
|
|
|
| 0.25% Metanil Yellow Solution |
|
0.25g |
|
Metanil yellow |
|
100.0ml |
|
Distilled water |
|
0.25ml |
|
Glacial acetic acid |
STAINING PROCEDURE:
- Treat all slides in Lugol's iodine for 10-60 minutes.
- Bleach in sodium bisulfite for 30 seconds.
- Rinse in distilled water for 2 minutes.
- Dehydrate in 70% ethyl alcohol for 1 minute.
- Stain in aldehyde fuchsin:
elastic fibers -- 5-10 minutes
islet beta cells -- 15-30 minutes
pituitary -- 30 minutes to 2 hours
NOTE: Periodically rinse in 70% ethyl alcohol and examine microscopically
for best results.
- Rinse in 2 changes of 70% ethyl alcohol.
- Counterstain as desired with the Azan method (2-3), the Van Gieson
method (2-12), or with metanil yellow for 5 minutes.
- Rinse in distilled water.
Blow dry.
Mount.
RESULTS:
Elastic fibers, mast cells, gastric chief cells, beta cells of pancreatic
islets, certain adenohypophyseal basophils, some (but not all) kinds
of mucin. Plastic background stain is intense, but should be regarded
as only a superficial nuisance as it does not affect the tissue components.
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. |
