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Visualization of Neurons Filled With Biotinylated-Lucifer Yellow Following Identification of Efferent Connectivity With Retrograde Transport

Visualization of Neurons Filled With Biotinylated-Lucifer Yellow Following Identification of Efferent Connectivity With Retrograde Transport

S J Hill, D L Oliver

J Neurosci Methods. 1993 Jan;46(1):59-68.

PMID: 8459723

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 in combination with neural connections and dendritic morphology.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP67769475	Lucifer Yellow CH dilithium salt		67769-47-5	Price

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