The delivery of genetic payloads to cells using genetic medicines is challenging to predict and the existing tools to assess delivery in animal models, although critical for discovering and advancing these drugs to the clinic, do not readily facilitate assessments of functional delivery throughout the entire body. To complement these existing techniques, our lab has used whole-body 3-D hyperspectral fluorescence cryo-imaging for the evaluation of functional delivery in whole animal specimens at high resolution. This instrument acquires hyperspectral fluorescence images of whole animal specimen while they are sectioned at micron-level resolution. In this study, mice were administered AAV9, a common adeno-associated viral delivery vehicle to express and image a fluorescent reporter. As the specimen is sectioned the acquisition can be paused to collect whole-body tissue samples which are then stained for immunofluorescence (IF) analysis. Herein, we describe a technique to reconstruct IF images into a single whole-body tissue specimen to be assessed alongside the co-registered cryo-images.
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