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

Here we provide a clearing-free protocol for processing intact, whole mount sub-cutaneous white adipose tissue (scWAT) for immunofluorescence as an alterna-tive to current clearing-based approaches. We use a combination of Z-depth reduction and autofluorescence quenching techniques to fluorescently label, im-age, and quantify adipose tissue innervation effectively throughout intact mouse tissues without the need for optical clearing or light sheet microscopy. This pro-tocol 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).

Automated summary

Here, we present a clearing-free protocol for processing intact sub-cutaneous white adipose tissue for immunofluorescence, which offers an alternative to current clearing-based approaches. This protocol utilizes Z-depth reduction and autofluorescence quenching techniques to effectively label and quantify adipose tissue innervation without the need for optical clearing or light sheet microscopy. It has been optimized and validated for adipose neurovascular labeling.