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In the present study, pancreas of rats were dissected and transferred to HEPES buffer (25 mM, pH 7.4). The control tissue pieces were kept in culture medium for one hour and the treated tissues were kept in same medium for 30 minutes and incubated with Insulin (10 nm and 100 nm) for another half hour, then tissues were transferred to Bouin's fixative (overnight at 40 ° Cc), cryosectioned (15 µm at -16 0 c) and subjected to immunocytochemical labeling with antibodies against Glucagon.

Glucagon Secreting Cells Responds to Insulin Secretion In vitro Using Immunocytochemistry