Journal article

COMBINED INTRACELLULAR INJECTION OF NEUROBIOTIN AND PREEMBEDDING IMMUNOCYTOCHEMISTRY USING SILVER-INTENSIFIED GOLD PROBES IN MYENTERIC NEURONS

HM YOUNG, WAA KUNZE, S POMPOLO, JB FURNESS, JC BORNSTEIN

JOURNAL OF NEUROSCIENCE METHODS | ELSEVIER SCIENCE BV | Published : 1994

Abstract

We have developed methods to examine the neurochemistry of synaptic inputs to individual myenteric neurons labeled by dye injection through intracellular recording electrodes. Myenteric neurons of the guinea-pig ileum were filled with Neurobiotin, fixed, washed in 50% ethanol, exposed to sodium cyanoborohydride, incubated in avidin-biotin-horseradish peroxidase and incubated in antisera to calretinin or calbindin. The Neurobiotin-filled cells were revealed using the diaminobenzidine (DAB) reaction. The tissue was examined at the light microscope level to determine the morphology and projections of the Neurobiotin-filled neurons, and then incubated in 1 nm gold-labeled secondary antibodies. F..

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