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Intracellular cholesterol visualization in brain tissue using D4H^∗.

ABSTRACT: Studying cholesterol biology in the brain has been greatly hindered by the lack of adequate cholesterol visualization techniques. Here, we present a protocol for using a high-affinity cholesterol probe D4H^∗-mCherry as a histology reagent in mouse or human brain tissue. We describe steps for D4H^∗ tissue treatment and crosslinking leading to stable labeling of intracellular membrane cholesterol. Furthermore, co-labeling with Rab5 endosomal marker and optimized buffers to reduce background enable punctate cholesterol visualization within the organelle membranes.

SUBMITTER: de Leeuw SM

PROVIDER: S-EPMC10762518 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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Intracellular cholesterol visualization in brain tissue using D4H∗.

de Leeuw Sherida M SM Nuriel Tal T

STAR protocols 20231213 1

Studying cholesterol biology in the brain has been greatly hindered by the lack of adequate cholesterol visualization techniques. Here, we present a protocol for using a high-affinity cholesterol probe D4H∗-mCherry as a histology reagent in mouse or human brain tissue. We describe steps for D4H∗ tissue treatment and crosslinking leading to stable labeling of intracellular membrane cholesterol. Furthermore, co-labeling with Rab5 endosomal marker and optimized buffers to redu ...[more]

PMID: 38100357

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