SeeDB: A simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction

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484 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)1154-1161
Number of pages8
JournalNature Neuroscience
Volume16
Issue number8
DOIs
Publication statusPublished - Aug 1 2013
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)

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