Rapid Haematoxylin and eosin (H&E) staining: Frozen section.

Frozen section

Frozen section cutting is a method by which it is possible to diagnose cancer very quickly. In case of any major operation, it is possible to diagnose the disease quickly by staining the section with suspicious tissue frozen section while keeping the patient in the operation theater. It usually takes 2 days to report by paraffin-sectioning. In that case the frozen section takes only 10 minutes.

2 types of microtomes are used for frozen section.

i) Freezing microtome using CO2 gas
ii) Refrigerated microtome (cryostat)

Make a slide by cutting a section on any one microtome.
With the section on the slide, one of the following 2 methods would have to be stained in the same way:

1. Rapid H&E

2. Toluidine blue

Rapid Haematoxylin and eosin (H&E) staining:

1. Keep slide with section in Haematoxvlin for one minute.
2. Wash in tap water. (rinse with tap water).
3. 1% Acid alcohol should be given one drip fast.
4. Wash in tap water. (rinse with tap water).
5. Ammonia vapor's should be taken quickly.
6. Rinse with tap water.
7. Put in 1% aqueous eosin for 3-6 seconds.
8. Rinse with tap water 

9. One drip at 95% alcohol
10. One drip in absolute alcohol
11. One drip in xylene
12. Mount in DPX
13. Examine under microscope

Toluidine blue staining:

1. Slide with section toluidine blue 0.5% at 30seconds to 1 minute should be submerged.
2. Rinse with water
3. Mount in water glycerin with cover slip.
4. Examine under microscope.