PMID: 8459723Jan 1, 1993Paper

Visualization of neurons filled with biotinylated-lucifer yellow following identification of efferent connectivity with retrograde transport

Journal of Neuroscience Methods
S J Hill, D L Oliver

Abstract

A new method is described that allows identification of neurons by retrograde transport, intracellular injection of biotin-labeled Lucifer Yellow, and a histochemical reaction of the biotin. Cells in the inferior colliculus that project to the ipsilateral thalamus are identified by injection of rhodamine-labeled latex beads into the medial geniculate body and retrograde transport in vivo. Several days later, the brains are fixed with aldehydes, and the inferior colliculus is cut on a vibratome. The rhodamine-labeled cells are observed with epi-fluorescent optics, impaled with intracellular pipettes under visual control, and injected with 9% Lucifer Yellow dilithium salt and 1% Lucifer Yellow cadaverine biotin-X. The biotinylated Lucifer Yellow is visualized by incubation of the slice overnight in avidin-biotin-HRP complex in the presence of 0.1% Triton X-100 at 4 degrees C. A diaminobenzidine reaction with simultaneous cobalt and nickel intensification follows. This method produces well-filled neuronal cell bodies, dendrites, and spines for light microscopic analysis. The non-fluorescent reaction product in these intracellularly filled cells may permit transmitter immunohistochemistry and synaptic fine structure to be studied i...Continue Reading

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Citations

Sep 18, 2002·Journal of Neuroscience Methods·Christopher J PaceSuzannah Bliss Tieman
Feb 12, 1999·Brain Research. Brain Research Protocols·G T Prusky, J E McKenna
Sep 9, 2011·Journal of Neurophysiology·Nancy F DayTeresa A Nick
Jul 26, 2011·Micron : the International Research and Review Journal for Microscopy·Shannon Modla, Kirk J Czymmek
Nov 9, 2000·Microscopy Research and Technique·D L Oliver
Nov 1, 1994·Microscopy Research and Technique·K L SmithJ W Swann
Apr 4, 1998·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·X J SunI A Meinertzhagen

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