Combined light and electron microscope immunocytochemical localization of scattered peptidergic neurons in the central nervous system
Article first published online: 3 FEB 2005
Copyright © 1986 Wiley-Liss, Inc.
American Journal of Anatomy
Volume 175, Issue 2-3, pages 179–195, February - March 1986
How to Cite
Anthony, E. L. P. and King, J. C. (1986), Combined light and electron microscope immunocytochemical localization of scattered peptidergic neurons in the central nervous system. Am. J. Anat., 175: 179–195. doi: 10.1002/aja.1001750207
- Issue published online: 3 FEB 2005
- Article first published online: 3 FEB 2005
- Manuscript Accepted: 5 OCT 1985
- Manuscript Received: 19 AUG 1985
A pre-embedding immunocytochemical technique is described for combined light and electron microscope study of peptidergic neurons in the central nervous system. The protocol is especially designed to overcome the sampling problems inherent in electron microscope study of structures, such as luteinizing hormone-releasing hormone (LHRH) neurons, that are scattered individually across large brain regions. The fixation methods outlined for several mammalian species include immersion and vascular perfusion with acrolein. Fine-structural preservation and LHRH immunoreactivity obtained with this fixative are compared to results with more conventional fixatives. Vibratome sectioning and a “pretreatment” regime, which prepare the tissues for immunocytochemistry, are described. Immunocytochemical labeling is done with free-floating sections and the peroxidase-antiperoxidase unlabeled antibody enzyme technique. Techniques are also described for the subsequent processing of immunoreacted sections for electron microscopy. These methods ensure that the processed sections are readily scanned by light microscopy, so that regions containing immunoreactive structures can be specifically chosen for electron microscope analysis. Sample electron micrographs are shown that illustrate some fine structural features of LHRH neurons in rats, bats, ferrets, and monkeys, as revealed with the techniques described.