Frozen Sections and Cytocentrifuge Preparations

Cryostat sections and cytocentrifuge preparations should be air-dried for at least 1 h but preferably overnight before immuno-staining. Before immunolabeling, cryostat sections should be fixed in cold acetone for 10 min and cytospin slides should be fixed for 90 s in a 1:1 mixture of acetone and methanol at room temperature. After fixation, follow the IGSS method for paraffin sections from step 3. Cytospin preparations are usually adequately covered by standard

Pancreatic Islet Imgages
Fig. 2. Photomicrograph of P-cells in a pancreatic islet of Langerhans-IGSS labeling of insulin (magnification x400).

25-^L aliquots of reagent. If immunolabeling is weak, the sections may be permeabilized for 5 min in 0.1% trypsin in TBS containing 0.1% calcium chloride at 37°C (16).

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