The stain, which I want to show, is a Trichrome overview stain according to Mallory. Frank Burr Mallory was an American Pathologist and Professor of Pathology at Harvard University. His staining protocol dates back to the year 1900. He describes a tissue fixation with Zenkerian fluid. My experience with Zenker is fine but even after fixation with Bouin and even after a simple formaldehyde fixation the color results are still fine although they are less powerful.
The protocol:
- Bring Coupes in Water
- Fuchsine acid 0.25% in AD, 5min
- Rinsing with AD
- Fixate and differentiate with phosphorus tungsten acid 5% in AD, 5min
- Rinsing with AD
- Aniline Blue/Orange G/Oxalic acid mixture, 7min
- Rinsing with AD
- Differentiate into two portions of ethanol 96% until no dye clouds are released
- Dehydrating
- Xylol and embedding in synthetic resin.
In most Histological books it is described that staining is done in specially designed staining cuvettes. However, my staining technique looks slightly different. In these cuvettes the dye consumption is quite high. Usually I only stain a few coupes at a time and the normal cup glasses are very suitable for that as well. These stains are done directly on the object glass and that works perfectly.
In this staining, two coupes are stained in a single process. Click on an image to view the series in large format. At the bottom of the image is the description of the work.
