A method for diagnosing POLYNEUROPATHY fine fibers in patients with celiac disease

摘要

1. A method for diagnosing polyneuropathy fine fibers in patients with celiac disease, which executes sampling three skin samples from the distal portion of the lower extremity (100 mm above the lateral malleolus), and then biopsies were fixed in a solution for fixing, and the solution as is used for fixing two drugs % solution lizinperiodat or paraformaldehyde-2% parafarmaldegid, picric acid (Zamboni solution) in 3 ml for 2 days at 4 ° C, after which the biopsies were washed for 10 minutes in a solution of sodium phosphate buffer 0.08 M, then incubated cryoprotective solution, which was prepared from 20 ml of glycerine, 2 ml of DMSO and 78 ml Sorensen solution at 4 ° C for 12 h, frozen biopsies were cut 50 microns, the solution was washed with sodium phosphate buffer 0.08 M 2 times 10 minutes, primary antibodies were added in the required dilution (1: 100, 1: 200 or 1: 500), incubated for 12 hours at room temperature in a humid chamber, washed biopsies solution of sodium phosphate buffer 0.08 M 2 times for 10 min, after which the secondary antibody at a dilution of 1: 100, is left on the table at room temperature for 1 h, the solution was washed with sodium phosphate buffer 0.08 M x ​​10 2 min, incubated in 30% methanol in sodium phosphate buffer for 30 min at room temperature, the solution was washed with biopsies sodium phosphate buffer 0.08 M 2 times for 5 min, then kept in biopsies AV mixture prepared based 1A: 1B: 100 PBS, in for 1 hour at room temperature, the solution was washed with sodium phosphate buffer 0.08 M 2 times for 10 minutes, after which the mixture was added to the biopsy sample prepared by mixing 5 ml