Biocytin Labeling

Biocytin labeling is a neuroanatomical method for tracing the structure and connections of individual neurons, helping researchers link cellular morphology with neural function. During intracellular recording, biocytin enters a neuron through the recording pipette and disperses throughout its soma, dendrites, and axon; after fixation, the biotin group is detected with labeled avidin or streptavidin, often producing a fluorescent or chromogenic signal. This approach enables detailed reconstruction of neuronal arbors, identification of axonal projections, and correlation of electrical properties with anatomy. Biocytin labeling is therefore valuable in circuit mapping, slice electrophysiology, developmental neuroscience, and studies of synaptic connectivity.

Biocytin Labeling - Related Videos

Research

JoVE EoE - Neuropathology

Labeling of Biocytin-Filled Interneurons in Rat Hippocampal Slices for Visualization

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2025

This video demonstrates the procedure for staining fixed rat brain slices containing biocytin-filled interneurons. The protocol includes incubation with avidin-biotin complex (ABC), staining with HRP and chromogenic substrate, preserving structures with osmium tetroxide, and embedding the tissue in resin for imaging.

Juxtasomal Biocytin Labeling to Study the Structure-function Relationship of Individual Cortical Neurons

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Cited by 22 •

2014

To understand the structure of neuronal networks, functional and morphological characterization of individual neurons is a necessity. Here, we demonstrate juxtasomal biocytin labeling, which allows electrophysiological recordings in the extracellular configuration, yet maintaining the ability to intracellularly label the neuron for post hoc reconstruction of dendritic and axonal architecture.

Immunostaining of Biocytin-filled and Processed Sections for Neurochemical Markers

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Cited by 28 •

2016

This protocol presents a method for the morphological recovery of neurons patched during electrophysiological recordings using biocytin filling and subsequent immunohistochemical postprocessing. We show that thick biocytin-filled sections that were stained and coverslipped can be restained with a second primary antibody days or months later.

Research

JoVE Journal - Neuroscience
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Biocytin Recovery and 3D Reconstructions of Filled Hippocampal CA2 Interneurons

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Cited by 4 •

2018

The protocol outlined here describes the immunofluorescence analysis, biocytin recovery and high-quality reconstructions of hippocampal CA2 interneurons following the intracellular electrophysiological recordings in vitro, allowing neuronal characterization and ultimately fine neuronal anatomy to be studied.

Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy

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Cited by 3 •

2025

This protocol presents a step-by-step guide for the Simultaneous Label-free Autofluorescence Multi-harmonic (SLAM) microscopic technique, including details on how to generate the laser light source, prepare a tissue sample, conduct imaging, and analyze the data. SLAM advances nonlinear microscopy by measuring four complementary label-free contrasts to investigate the tissue microenvironment.

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