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Journal of Visualized Experiments
Article . 2020 . Peer-reviewed
Data sources: Crossref
Journal of Visualized Experiments
Article . 2020 . Peer-reviewed
Data sources: Crossref
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A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Authors: Kristin N, Kirchner; Hailong, Li; Adam R, Denton; Steven B, Harrod; Charles F, Mactutus; Rosemarie M, Booze;

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Abstract

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered tissue samples. Traditional immunolabeling procedures require cutting the sample into thin sections, which restricts the ability to label and examine intact structures. However, if brain tissue can remain intact during processing, structures and circuits can remain intact for the analysis. Previously established clearing methods take significant time to completely clear the tissue, and the harsh chemicals can often damage sensitive antibodies. The iDISCO method quickly and completely clears tissue, is compatible with many antibodies, and requires no special lab equipment. This technique was initially validated for the use in mice tissue, but the current protocol adapts this method to image hemispheres of control and transgenic rat brains. In addition to this, the present protocol also makes several adjustments to preexisting protocol to provide clearer images with less background staining. Antibodies for Iba-1 and tyrosine hydroxylase were validated in the HIV-1 transgenic rat and in F344/N control rats using the present hydrophobic tissue clearing method. The brain is an interwoven network, where structures work together more often than separately of one another. Analyzing the brain as a whole system as opposed to a combination of individual pieces is the greatest benefit of this whole brain clearing method.

Keywords

Cholera Toxin, Dehydration, Staining and Labeling, Tyrosine 3-Monooxygenase, Dopaminergic Neurons, Calcium-Binding Proteins, Microfilament Proteins, Brain, Microtomy, Immunohistochemistry, Antibodies, Rats, Inbred F344, Rats, Mice, Animals, Microglia, Hydrophobic and Hydrophilic Interactions

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selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
1
Average
Average
Average
bronze