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Functional Calcium Imaging in Developing Cortical Networks
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Fast Calcium Imaging with Optical Sectioning via HiLo Microscopy.

Marcel A Lauterbach1, Emiliano Ronzitti1, Jenna R Sternberg2

  • 1Wavefront-Engineering Microscopy Group, Neurophotonics Laboratory, CNRS UMR8250, University Paris Descartes, Sorbonne Paris Cité, Paris, France.

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Summary

HiLo microscopy offers cost-effective, high-speed optical sectioning for in vivo calcium imaging. This technique enables detailed visualization of neuronal activity deep within tissues, overcoming limitations of traditional methods.

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Area of Science:

  • Neuroscience
  • Biophysics
  • Optical Microscopy

Background:

  • Calcium imaging is crucial for non-invasive neuronal activity monitoring.
  • High-speed optical sectioning is needed for deep tissue imaging but is challenging and costly.
  • Existing methods struggle to balance resolution, speed, and cost for in vivo applications.

Purpose of the Study:

  • To develop an accessible and cost-effective solution for high-speed in vivo optical sectioning.
  • To demonstrate the efficacy of HiLo microscopy for calcium imaging in dense neuronal populations.
  • To compare HiLo microscopy with wide-field microscopy for calcium imaging and 3D reconstruction.

Main Methods:

  • Implementation of HiLo microscopy for in vivo calcium imaging.
  • Acquisition of functional calcium imaging data at 100 frames per second.
  • Quantification of optical sectioning capabilities and background fluorescence reduction.

Main Results:

  • Successful recording of large calcium signals from dense neuronal populations at high acquisition rates.
  • Demonstrated superior optical sectioning and 3D reconstruction capabilities compared to wide-field microscopy.
  • Discriminated neuronal activity in zebrafish spinal cord motor neurons at different depths, observing distinct somatic and axonal signal dynamics.

Conclusions:

  • HiLo microscopy provides robust, high-frame-rate optical sectioning for in vivo calcium imaging at low cost.
  • This method effectively visualizes neuronal activity deep within tissues, including motor neurons in zebrafish embryos.
  • The setup is adaptable to existing microscopes, offering broad applicability for neuroscience research.