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[Music] load the paraffin-embedded tissue onto a microtome and cut into five micrometer sections [Music] transfer the tissue to a bath of distilled water at 40 degrees Celsius and mount on a glass slide suitable for immunostaining dry slides overnight at 56 degrees Celsius follow the slide treatment steps as listed here xylene solution 3 times 5 minutes each 100 percent ethanol 3 times 5 minutes each 95 percent ethanol for 3 minutes 70 percent ethanol for 3 minutes ro/di reverse osmosis and deionization and water 3 times 1 minute each [Music] for chroma genic staining x block endogenous peroxidase activity by incubating slides in 3% hydrogen peroxide solution in methanol at room temperature for 15 minutes perform antigen retrieval using a microwave or a pressure cooker antigen retrieval with a microwave prepare a 100 mil solution of 1x citrate buffer by mixing 10 mils of bio legends 10x citrate buffer catalog number 4 2 0 9 0 1 with 90 mils of distilled water [Music] microwaved tissue