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MIRAVA POLYSCOPE – All in one and on for all: the perfect image
Science beyond Barriers

abberior instruments

Neurobiology, Physiology

2018
Nature communications

Quantitative optical nanophysiology of Ca 2+ signaling at inner hair cell active zones

Authors:

Neef, J., Urban, N. T., Ohn, T. L., Frank, T., Jean, P., Hell, S. W., ... & Moser, T.

Keywords:

Ca2+ imaging, Hair cell, Synaptic transmission

Abstract:

Ca2+ influx triggers the release of synaptic vesicles at the presynaptic active zone (AZ). A quantitative characterization of presynaptic Ca2+ signaling is critical for understanding synaptic transmission. However, this has remained challenging to establish at the required resolution. Here, we employ confocal and stimulated emission depletion (STED) microscopy to quantify the number (20–330) and arrangement (mostly linear 70 nm × 100–600 nm clusters) of Ca2+ channels at AZs of mouse cochlear inner hair cells (IHCs). Establishing STED Ca2+ imaging, we analyze presynaptic Ca2+ signals at the nanometer scale and find confined elongated Ca2+ domains at normal IHC AZs, whereas Ca2+ domains are spatially spread out at the AZs of bassoon-deficient IHCs. Performing 2D-STED fluorescence lifetime analysis, we arrive at estimates of the Ca2+ concentrations at stimulated IHC AZs of on average 25 µM. We propose that IHCs form bassoon-dependent presynaptic Ca2+-channel clusters of similar density but scalable length, thereby varying the number of Ca2+ channels amongst individual AZs.

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Superresolution & Confocal Systems

  • Overview
  • MINFLUX
  • MIRAVA POLYSCOPE
  • INFINITY
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  • STEDYCON
  • Software Overview
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Superresolution & Confocal Modules

  • Overview
  • MINFLUX Module
  • MATRIX Detector
  • TIMEBOW Imaging
  • FLEXPOSURE Illumination
  • RAYSHAPE Mirror
  • TRUESHARP Deconvolution
  • EASY3D
  • RAINBOW Detection
  • STED Lasers
  • Autoalignment
  • Autofocus
  • Excitation Lasers
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