AbstractThis unit describes a high‐resolution [3H]2‐deoxyglucose (2DG)/immunohistochemistry double labeling protocol which enables simultaneous visualization of metabolic and immunohistochemical markers (e.g., neurotransmitter‐specific epitopes) in the same tissue section. This approach generates single‐cell metabolic measures from large (many thousands) samples of neurons, and allows the assignment of putative neurotransmitter types to large samples of functionally assayed single neurons. Thus, it provides a unique opportunity to assay the metabolic activities of histochemically identified neural elements throughout an entire pathway. [3H]2DG injection and perfusion fixation of hamsters or mice is described first, followed by immunohistochemical staining of brain slices for a monoclonal antibody against glutamic acid decarboxylase‐6 (GAD‐6). Included are the steps for dipping and developing slides for autoradiography. An alternate protocol gives details of staining for cytochrome oxidase (CO).
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