|
ZENKER'S FLUID/FIXATIVE
| Potassium dichromate |
2.5 g
|
| Mercuric chloride |
6.0 g
|
| Distilled water |
100 ml
|
| Acetic acid, glacial (add just before use) |
5.0 ml
|
Dissolve the potassium dichromate and mercuric chloride in the water while
heating. Use small pieces of tissue: 2 cm square or rectangle, not more
than 2-4 mm thick). Fix 6-8 hours; longer times will make tissue brittle
and difficult to section. Wash off excess fixative with running water
at least l~ hours. After fixation and washing, tissue may be stored in
70% EtOH. Remove mercuric chloride crystals before staining by immersing
the slides in alcoholic iodine after the paraffin has been removed.
How to remove mercuric chloride crystals:
- Deparaffinize slides through xylene, 100% and 95%
EtOH.
- Place in alcoholic iodine solution ( 0.5% iodine in
80% EtOH) for 5-10 minutes.
- Wash in water or rinse in 80% EtOH.
- Place in a solution of 5% sodium thiosulfate (hypo)
for 2-5 minutes to bleach out the iodine.
- Wash thoroughly in running tap water (5 minutes) to
remove the hypo.
- Stain
|