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Jake Willows, Kristy Townsend, Magdalena Blaszkiewicz 2021. Adipose depot innervation: whole mount staining, imaging, quantification. protocols.io dx.doi.org/10.17504/protocols.io.brs2m6geCopy Citation Copied
URL: https://dx.doi.org/10.17504/protocols.io.brs2m6ge
Authors: Jake Willows, Kristy Townsend, Magdalena Blaszkiewicz
Group: Townsend Lab Neurobiology & Energy Balance
Summary: AbstractLittle is known about the diversity and function of adipose tissue nerves due, in part, to the inability to effectively visualize the various nerve subtypes residing within these tissues. The tools currently available for researchers to image and quantify adipose tissue innervation are limited and dependent on optical clearing techniques and light sheet microscopy. Here we present a method of tissue processing that uses a method of mechanically compressing tissue to decrease tissue thickness in the z-axis by expanding it in the x and y-axes whilst leaving cells intact. This has been combined with autofluorescence quenching techniques to permit imaging of intact whole tissues on both widefield and confocal microscopes and a complementary means to perform whole tissue neurite density quantification. We have included examples of how this technique can be used to further our current knowledge of adipose-nerve communication by characterizing the nerves, nerve-subtypes, and neurovascular interactions within the inguinal subcutaneous white adipose tissue in mice.
Affiliations: Ohio State University, Columbus, Ohio State University, Ohio State University, Columbus
Version: 2
Publication Date: 2021
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Source: Protocols.io