摘要：SummaryLittle is known about the diversity and function of adipose tissue nerves, due in part to the inability to effectively visualize the tissue’s diverse nerve subtypes and the patterns of innervation across an intact depot. The tools to image and quantify adipose tissue innervation are currently limited. Here, we present a method of tissue processing that decreases tissue thickness in the z-axis while leaving cells intact for subsequent immunostaining. This was combined with autofluorescence quenching techniques to permit intact whole tissues to be mounted on slides and imaged by confocal microscopy, with a complementary means to perform whole tissue neurite density quantification after capture of tiled z-stack images. Additionally, we demonstrate how to visualize nerve terminals (the neuro-adipose nexus) in intact blocks of adipose tissue without z-depth reduction. We have included examples of data demonstrating nerve subtypes, neurovascular interactions, label-free imaging of collagen, and nerve bundle digital cross-sections.Graphical abstractDisplay OmittedHighlights•Whole depot adipose tissue innervation was imaged and quantified by a novel method•Numerous aspects of adipose nerve heterogeneity were observed by microscopy•We have identified a nerve terminal in adipose, the neuro-adipose nexusCell biology; Functional aspects of cell biology; Methodology in biological sciences
