Fig. 1

PEGASOS recirculation procedure achieves whole-body transparency and enables whole-head imaging with no blind area. a Brief description of the PEGASOS recirculation procedure. b–c Adult CAG-EGFP mouse (2-months old) was imaged before (b) and after (c) the recirculation procedure. The skin, eyeballs and tongue were removed to facilitate penetration. Other tissues including brain, bones, teeth, muscles, and glands remained intact. d–k Body parts and internal organs were dissected after clearing. l–q After clearing, the whole head was imaged with a two-photon microscope in both ventral-to-dorsal (V–D) and dorsal-to-ventral (D–V) directions. The two image stacks were stitched together with Image J. o–q Show optical sections acquired at different depths in the D–V direction. SHG second harmonic generation signal. Scale bars in d–q, 5 mm