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A clearing-free protocol for imaging intact whole adipose tissue innervation in mice

Jake W. Willows, Magdalena Blaszkiewicz, Kristy L. Townsend

2022STAR Protocols16 citationsDOIOpen Access PDF

Abstract

Here we provide a clearing-free protocol for processing intact, whole mount subcutaneous white adipose tissue (scWAT) for immunofluorescence as an alternative to current clearing-based approaches. We use a combination of Z-depth reduction and autofluorescence quenching techniques to fluorescently label, image, and quantify adipose tissue innervation effectively throughout intact mouse tissues without the need for optical clearing or light sheet microscopy. This protocol has been optimized and validated for adipose neurovascular labeling. For complete details on the use and execution of this protocol, please refer to Willows et al. (2021).

Topics & Concepts

Adipose tissueAutofluorescenceImmunofluorescenceClearingWhite adipose tissueSubcutaneous adipose tissueNeurovascular bundleProtocol (science)Biomedical engineeringPathologyMedicineFluorescenceInternal medicineOpticsImmunologyEconomicsAlternative medicineFinancePhysicsAntibodyAdipose Tissue and MetabolismAdvanced Fluorescence Microscopy TechniquesSingle-cell and spatial transcriptomics
A clearing-free protocol for imaging intact whole adipose tissue innervation in mice | Litcius