Teased Fiber Preparation of Myelinated Nerve Fibers from Peripheral Nerves for Vital Dye Staining and Immunofluorescence Analysis
Glial cells regulate a wide variety of neuronal functions during physiological and pathological conditions. Therefore, the study of glial cells and their association with axons is of paramount importance in order to understand the physiology of the nervous system. This chapter describes a detailed protocol to prepare and stain teased nerve fibers from peripheral nerves using fluorescent indirect immunolabeling and staining with vital dyes. For immunofluorescence analysis, we describe techniques to study the axonal compartment and the expression of cytoplasmic and plasma membrane proteins in Schwann cells.
Key wordsPeripheral nervous system Schwann cell Nerve fiber Immunofluorescence Teased fibers
We wish to thank all the members of the Court Lab for their contributions to this protocol. This work was supported by the Center for Integrative Biology, Universidad Mayor, FONDECYT-1150766, Geroscience Center for Brain Health and Metabolism (FONDAP-15150012), Ring Initiative ACT1109, and Canada-Israel Health Research initiative, jointly funded by the Canadian Institutes of Health Research; the Israel Science Foundation; the International Development Research Centre, Canada; and the Azrieli Foundation, Canada.