三色染色有时候用ZENKER固定剂或含有重铬酸钾液体做媒染剂。还有一种六氨银染色用铬酸处理。你是做三色染色还是做别的什么染色。
下面是重铬酸钾在染色中的应用参考：
http://members.pgonline.com/~bryand/StainsFile/stain/conektv/tri_milligan.htm
有关铬酸在染色中的应用
Grocott's Silver-Methenamine Method for Fungus
Fixation: 10% Neutral buffered formalin
Technique: Paraffin sections cut at 4 microns
Control: Tissue containing fungus
Solutions: 5% aqueous silver nitrate, with 3% methenamine
10% Chromic Acid Solution
Chromium trioxide 5.0 g
Distilled water 50.0 ml
1% Sodium Bisulfite Solution
Sodium bisulfite 0.5 g
Distilled water 50.0 ml
2% Sodium Thiosulfate
Sodium thiosulfate 2.5 g
Distilled water 50.0 ml
Methenamine-Silver Nitrate Solution (Stock)
5% Silver nitrate 5.0 ml
3% Methenamine 100.0 ml
A white precipitate forms but immediately dissolves on shaking. Clear solutions remain usable for months. Store in refrigerator.
Methenamine-Silver Nitrate Solution (working)
Methenamine-silver nitrate solution (stock) 25.0 ml
Distilled water 25.0 ml
5% Borax 2.0 ml
Make this solution fresh each time.
0.1% Gold Chloride Solution
1% Gold Chloride solution 5.0 ml
Distilled water 45.0 ml
This solution may be used until brown precipitate appears and the solution is cloudy.
1% Light Green Solution (stock)
Light green SF yellowish 1.0 g
Distilled water 100.0 ml
Glacial acetic acid 0.2 ml
Thymol 2.0 grains
1% Light Green Solution (working)
Light green, stock 10.0 ml
Distilled water 40.0 ml
Staining Procedure:
Attach to automatic stainer
Deparaffinize sections and hydrate to distilled water. Slides previously stained with most other stains may be used by removing coverslips and running through xylene and alcohols to water. Subsequent chromic acid treatment will remove any remaining stain.
Oxidize in fresh 10% chromic acid solution for 10 minutes at room temperature. (Note: this step may also be performed in a vented laboratory microwave set at 60 C for 90 seconds).
Rinse in distilled water for a few seconds.
Rinse until clear in 1% sodium bisulfite to remove any residual chromic acid.
Rinse in distilled water 5 times.
Place slides in Silver Methenamine Solution and microwave for 45 seconds. Let slides set for 2 minutes. Examine microscopically. Fungi should be dark brown. If staining is not complete microwave again in the hot working solution for 10 seconds. Re-examine slides. Repeat this step until the desired staining intensity is reached.
Rinse in 5 changes of distilled water.
Tone in 0.1% gold chloride solution until sections are gray-lavender, 1 minute.
Rinse in distilled water
Remove unreduced silver with 2% thiosulfate (hypo) for 1 minute.
Wash thoroughly in tap water.
Counterstain with working light green solution for 15 seconds.
Dehydrate, clear and mount with xylene soluble media.
Results:
Fungi - sharply delineated black
Mucin - taupe to dark gray
Inner parts of mycelia & hyphae - old rose
Background - pale green
Adapted for the Jefferson Regional Medical Center Histo-Path Laboratory
References
Grocott, R.G.: American Journal of Clinical Pathology, 25: pp 975-979, 1955.
