title: Optical clearing of fixed brain samples using SeeDB.
abstract: Large-scale three-dimensional fluorescence imaging is essential for comprehensive and quantitative understanding of neuronal circuitry. We describe a water-based optical clearing agent, SeeDB, which clears fixed brain samples in a few days leaving many types of fluorescent dyes unquenched, including fluorescent proteins and lipophilic neuronal tracers. This method maintains a constant sample volume during the clearing procedure, an important factor to keep cellular morphology intact. After optical clearing with SeeDB, we can reach a depth of >1000 µm with confocal microscopy. When combined with two-photon microscopy, SeeDB allows us to image fixed mouse brains at millimeter-scale level.
authors: Laboratory for Sensory Circuit Formation, RIKEN Center for Developmental Biology, Kobe, Japan; Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
authors: Meng-Tsen
authors: MT
authors: Ke
authors: Takeshi
authors: T
authors: Imai
mesh_terms: D000818:Animals; D001921:Brain / Q000196:embryology*; D001923:Brain Chemistry / Q000502:physiology; D005404:Fixatives / Q000032:analysis; D005632:Fructose / Q000032:analysis*; D005990:Glycerol / Q000031:analogs & derivatives* / Q000032:analysis; D051379:Mice; D008822:Mice, Transgenic; D036641:Microscopy, Fluorescence, Multiphoton / Q000379:methods*; D012996:Solutions / Q000032:analysis*
chemical_list: D005404:Fixatives; C583792:SeeDB solution; D012996:Solutions; D005632:Fructose; D005990:Glycerol
keywords: fluorescent imaging; optical clearing; two-photon microscopy