A clearing-free protocol for imaging intact whole adipose tissue innervation in mice
Jake W. Willows, Magdalena Blaszkiewicz, Kristy L. Townsend
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