2-12. Van Gieson Stain (Ref 8)
Staining Procedures For Plastic Embedded Tissue
Verified at the Applications Laboratory of the Biomedical Division, Sorvall
Microtomes
SOLUTIONS:
| Celestin Blue Solution |
|
0.5g |
|
Celestin blue |
|
5.0g |
|
Ammonium ferric sulfate |
|
100.0ml |
|
Distilled water
Filter before use. |
|
|
|
|
| Weigert's Hematoxylin |
|
|
|
See formula in procedure 2-7,
c |
|
|
|
|
| Van Gieson's Solution |
|
20.0ml |
|
Acid fuchsin, 1% |
|
25.0ml |
|
Picric acid, saturated |
STAINING PROCEDURE:
- Treat in celestin blue for 10 minutes.
- Rinse in distilled water.
- Stain in Weigert's hematoxylin for 10 minutes.
- Rinse in tap water.
- Differentiate in acid alcohol for 2 to 3 quick dips.
- Wash in tap water very briefly.
- Dip in ammonia water until sections are bright blue; 2-3 dips.
- Wash in running tap water for 10-20 minutes.
- Stain in Van Gieson's solution for 15 minutes or longer.
- Wash in distilled water.
- Blow dry.
- Mount.Solutions:
RESULTS:
Nuclei stain -- black; muscle -- yellow; collagen -- red. Very thin
sections are often pale since less material is available to accept
stain.
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. |
