Super-Resolution Mapping of Neuronal Circuitry With an Index-Optimized Clearing Agent

Meng Tsen Ke, Yasuhiro Nakai, Satoshi Fujimoto, Rie Takayama, Shuhei Yoshida, Tomoya S. Kitajima, Makoto Sato, Takeshi Imai

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

Super-resolution imaging deep inside tissues has been challenging, as it is extremely sensitive to light scattering and spherical aberrations. Here, we report an optimized optical clearing agent for high-resolution fluorescence imaging (SeeDB2). SeeDB2 matches the refractive indices of fixed tissues to that of immersion oil (1.518), thus minimizing both light scattering and spherical aberrations. During the clearing process, fine morphology and fluorescent proteins were highly preserved. SeeDB2 enabled super-resolution microscopy of various tissue samples up to a depth of >100 μm, an order of magnitude deeper than previously possible under standard mounting conditions. Using this approach, we demonstrate accumulation of inhibitory synapses on spine heads in NMDA-receptor-deficient neurons. In the fly medulla, we found unexpected heterogeneity in axon bouton orientations among Mi1 neurons, a part of the motion detection circuitry. Thus, volumetric super-resolution microscopy of cleared tissues is a powerful strategy in connectomic studies at synaptic levels.

Original languageEnglish
Pages (from-to)2718-2732
Number of pages15
JournalCell Reports
Volume14
Issue number11
DOIs
Publication statusPublished - Mar 22 2016

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Tissue
Aberrations
Light scattering
Neurons
Microscopy
Microscopic examination
Connectome
Imaging techniques
Light
Refractometry
Optical Imaging
Immersion
N-Methyl-D-Aspartate Receptors
Mountings
Diptera
Synapses
Axons
Refractive index
Oils
Spine

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Super-Resolution Mapping of Neuronal Circuitry With an Index-Optimized Clearing Agent. / Ke, Meng Tsen; Nakai, Yasuhiro; Fujimoto, Satoshi; Takayama, Rie; Yoshida, Shuhei; Kitajima, Tomoya S.; Sato, Makoto; Imai, Takeshi.

In: Cell Reports, Vol. 14, No. 11, 22.03.2016, p. 2718-2732.

Research output: Contribution to journalArticle

Ke, Meng Tsen ; Nakai, Yasuhiro ; Fujimoto, Satoshi ; Takayama, Rie ; Yoshida, Shuhei ; Kitajima, Tomoya S. ; Sato, Makoto ; Imai, Takeshi. / Super-Resolution Mapping of Neuronal Circuitry With an Index-Optimized Clearing Agent. In: Cell Reports. 2016 ; Vol. 14, No. 11. pp. 2718-2732.
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