Vybrant Dyes -- Useful For Marking Electrode Tract

May 17, 2017

BlueskyReddit
Albert Einstein College of Medicine
Dominick P. Purpura Department of Neuroscience
Graduate student

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Company:

Invitrogen

Product Name:

Vybrant® Multicolor Cell-Labeling Kit (DiO, DiI, DiD Solutions, 1 mL each)

Catalog Number:

V22889

Our laboratory uses in vitro and in vivo electrophysiology and calcium imaging to understand how the cerebellum interacts with the rest of the brain. Vybrant dyes are extremely useful to us for labeling electrode tracts after acute, in vivo electrophysiological recordings or for labeling the recording sites of implanted electrodes. These dyes enable post-hoc identification of recording sites.

Experimental Design and Results Summary

Application

In vivo electrophysiology

Starting Material

Live mouse

Protocol Overview

It is useful to transfer 200 ul of dye into a small PCR tube. Coat electrode (or implant) 8-10 times with dye by dipping it into the dye in the tube. Let it dry 15-30 seconds in between each coating. Let it dry for 20 minutes. For best results, repeat this procedure 3 times. After experiment, perfuse animal and cryopreserve and section the brain. Image sections under a standard fluorescent microscope.

Tips

It is VERY important that the dye is given time to dry. If recording from deeper parts of the brain, make sure the tip of electrode is coated very well.

Results Summary

Coating electrodes or implants with these dyes before the experiment enables post-hoc location of recording sites. The dyes are very strong and the entire electrode tract can usually be visualized under a standard fluorescent microscope.

DOI or PMID #

N/A

Additional Notes

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Summary

The Good

Extremely bright dye enables exact identification of recording sites after in vivo electrophysiological experiments.

The Bad

It requires time for the dye to dry completely. It will not work well if dye does not dry.

The Bottom Line

These dyes are extremely useful for labeling electrode tracts in vivo. Especially if your target recording sites are small, these dyes are essential for making sure that you hit your target. It is worth both the time and expense.

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