title: SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction.
abstract: We report a water-based optical clearing agent, SeeDB, which clears fixed brain samples in a few days without quenching many types of fluorescent dyes, including fluorescent proteins and lipophilic neuronal tracers. Our method maintained a constant sample volume during the clearing procedure, an important factor for keeping cellular morphology intact, and facilitated the quantitative reconstruction of neuronal circuits. Combined with two-photon microscopy and an optimized objective lens, we were able to image the mouse brain from the dorsal to the ventral side. We used SeeDB to describe the near-complete wiring diagram of sister mitral cells associated with a common glomerulus in the mouse olfactory bulb. We found the diversity of dendrite wiring patterns among sister mitral cells, and our results provide an anatomical basis for non-redundant odor coding by these neurons. Our simple and efficient method is useful for imaging intact morphological architecture at large scales in both the adult and developing brains.
authors.affiliation: Laboratory for Sensory Circuit Formation, RIKEN Center for Developmental Biology, Kobe, Japan.
authors.forename: Meng-Tsen
authors.initials: MT
authors.lastname: Ke
authors.forename: Satoshi
authors.initials: S
authors.lastname: Fujimoto
authors.forename: Takeshi
authors.initials: T
authors.lastname: Imai
mesh_terms: D000818:Animals; D001565:Benzoates / Q000494:pharmacology; D019905:Benzyl Alcohol / Q000494:pharmacology; D001921:Brain / Q000187:drug effects / Q000196:embryology / Q000648:ultrastructure*; D003712:Dendrites / Q000648:ultrastructure; D018274:Electroporation; D005260:Female; D005404:Fixatives / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000032:analysis* / Q000737:chemistry; D005557:Formaldehyde / Q000494:pharmacology; D005632:Fructose / Q000737:chemistry / Q000494:pharmacology*; D055879:Gene Knock-In Techniques; D005990:Glycerol / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D016591:Histocytological Preparation Techniques; D008027:Light; D051379:Mice; D008810:Mice, Inbred C57BL; D008813:Mice, Inbred ICR; D008822:Mice, Transgenic; D008853:Microscopy / Q000379:methods; D009415:Nerve Net / Q000648:ultrastructure*; D009474:Neurons / Q000648:ultrastructure; D009812:Odorants; D009830:Olfactory Bulb / Q000166:cytology / Q000187:drug effects; D055696:Olfactory Perception / Q000502:physiology; D010647:Phenyl Ethers / Q000494:pharmacology; D011108:Polymers / Q000494:pharmacology; D011247:Pregnancy; D012542:Scattering, Radiation; D012996:Solutions / Q000737:chemistry / Q000494:pharmacology*; D013395:Sucrose / Q000494:pharmacology; D016707:Tissue Fixation / Q000379:methods; D014508:Urea / Q000494:pharmacology
chemical_list: D001565:Benzoates; D005404:Fixatives; D005456:Fluorescent Dyes; D010647:Phenyl Ethers; D011108:Polymers; C568745:ScaleA2 solution; C583792:SeeDB solution; D012996:Solutions; D005557:Formaldehyde; C076624:dibenzyl ether; D005632:Fructose; D013395:Sucrose; D014508:Urea; D019905:Benzyl Alcohol; C006723:benzyl benzoate; D005990:Glycerol; C003043:paraform