Immunostaining of Interneurons in a Rat Hippocampal Section

In dit artikel

Samenvatting

Swietek, B., et al., Immunostaining of Biocytin-filled and Processed Sections for Neurochemical Markers. J. Vis. Exp. (2016)

This video demonstrates the process of visualizing a biocytin-filled interneuron in a rat hippocampal section, highlighting its neurochemical identity through the use of specific antibodies and fluorescence microscopy.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Staining of the Primary Antibody and Biocytin

(Day 1)
NOTE: The following immunostaining procedure is for free-floating sections and requires continuous shaking at a low speed (2 revolutions/min, rpm) on a shaker for all incubation steps.

Wash the tissue 3x for 10 min each in 0.1 M PBS (pH = 7.4).
NOTE: 0.1 M PBS can be commercially purchased or made in the laboratory as follows (makes 1 L): 8 g of NaCl, 0.2 g of....

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Resultaten

Biocytin-labeled neuron, fluorescence microscopy, 3D orientation diagram with XYZ axes, 50 µm scale.
Figure 1. Suggested Plane for Approaching Cells for Biocytin Fills. The image illustrates a Granule Cell (GC) filled with biocytin during recording that has been processed to reveal biocytin (in red using 594-conjugated streptavidin). The GC has it.......