Acetylcholine receptor display cytochemistry experiments

Summary

Acetylcholine receptors are distributed at neuromuscular junctions, etc. Snake toxin binds tightly to them and connects to peroxidase, and the distribution of acetylcholine receptors can be observed under the electron microscope by chemical reactions. Source: Immunohistochemistry Experimental Techniques and Applications

Operation method

basic program

Materials and Instruments

Tissue samples
Potassium phosphate Glutaraldehyde KOH Sucrose Potassium pyroantimonate Osmium acid

Move

1. The tissue is cut into small pieces of about 1 mm³ and fixed with 0.09 mol/L potassium phosphate (or potassium oxalate)-3% glutaraldehyde (pH 7.3, adjust the pH with 0.1%-1% KOH) for more than 4 h at 4℃. Microwave irradiation is also recommended at the early stage of fixation to accelerate the penetration of the fixative into the cells (for the use of microwave irradiation, see the relevant section). 2.

2. After fixation, rinse with 0.09 mol/L potassium phosphate (or potassium oxalate)-4% sucrose (pH 7.3) for 2-4 h, and change the solution 3 times. 3. 1.5% to 2.5% of the cellular fluid should be rinsed with potassium phosphate (or potassium oxalate)-4% sucrose (pH 7.3).

3. 1.5%~2% potassium pyroantimonate-1% osmiic acid fixed at 4℃ for 2 h. 4. pH 10 distilled water.

4. pH 10 distilled water rinsing 3 times × 10 min, in order to wash out the residual pyroantimonate.

5. Dewatering, embedding as usual, sectioning, staining or no staining.

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