2-4. Modified Nocht's Azure-Eosin (Refs
1, 2)
Staining Procedures For Plastic Embedded Tissue
Verified at the Applications Laboratory of the Biomedical Division, Sorvall
Microtomes
SOLUTIONS:
| 0.1% Azure A |
|
0.2g |
|
Azure A |
|
200.0ml |
|
Distilled water |
|
|
|
|
| 0.1% Eosin B |
|
0.2g |
|
Eosin B |
|
200.0ml |
|
Distilled water |
|
|
|
|
| 0.1 M Citric Acid |
|
21.0g |
|
Citric acid |
|
qs to 1000.0ml |
|
Distilled water |
|
|
|
|
| 0.2 M Disodium Phosphate |
|
28.4g |
|
Disodium phosphate |
|
qs to 1000.0ml |
|
Distilled water |
|
|
|
|
| Working Solution |
|
25.0ml |
|
Distilled water |
|
8.0ml |
|
Azure A, 0.1% |
|
8.0ml |
|
Eosin B, 0.1% |
|
1.2ml |
|
Citric acid, 0.1 M
Disodium phosphate to make pH 4.8 to 5.2 (approx. 0.8ml). Lower pH levels
give redder results, higher levels more blue. |
|
5.0ml |
|
Acetone |
|
|
|
Filter. Solution stable for 1-2 weeks. May need refiltering
each week. |
STAINING PROCEDURE:
- Stain approximately 1 hour in working solution. (Overstaining
is not possible.)
Blow dry.
Mount.
RESULTS:
Cell granules, microorganisms, nuclei: similar to Giemsa
(2-6).
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. |
