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METHODS article

Front. Neurosci.
Sec. Neuroscience Methods and Techniques
Volume 18 - 2024 | doi: 10.3389/fnins.2024.1445020

Imaging Distinct Neuronal Populations with a Dual Channel miniScope

Provisionally accepted
Giovanni Barbera Giovanni Barbera 1,2*Rashmi Thapa Rashmi Thapa 3Navin Adhikari Navin Adhikari 3Yun Li Yun Li 3Da-Ting Lin Da-Ting Lin 2,4*
  • 1 National Institutes of Health (NIH), Bethesda, United States
  • 2 Behavioral Neuroscience Research Branch, Neural Engineering Section, Intramural Research Program (NIDA), Baltimore, Maryland, United States
  • 3 Department of Zoology and Physiology, University of Wyoming, Laramie, Wyoming, United States
  • 4 The Solomon H Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland, United States

The final, formatted version of the article will be published soon.

    Miniature fluorescence microscopes (miniscopes) are one of the most powerful and versatile tools for recording large scale neural activity in freely moving rodents with single cell resolution. Recent advances in the design of genetically encoded calcium indicators (GECIs) allow to target distinct neuronal populations with non-overlapping emission spectral profiles. However, conventional miniscopes are limited to a single excitation, single focal plane imaging, which does not allow to compensate for chromatic aberration and image from two spectrally distinct calcium indicators. In this paper we present an open-source dual channel miniscope capable of simultaneous imaging of genetically or functionally distinct neuronal populations. Chromatic aberrations are corrected using an electrowetting lens (EWL), which allows fast focal plane change between frames. To demonstrate the capabilities of the dual channel miniscope, we labeled layer specific excitatory neurons or inhibitory interneurons in the medial prefrontal cortex (mPFC) with a red fluorescence protein, and simultaneously imaged neural activity of distinct neuronal populations of freely moving mice via a green GECI.

    Keywords: Miniscope, calcium imaging, dual color imaging, miniature fluorescence microscope, liquid lens, EWL, single-photon, open source

    Received: 06 Jun 2024; Accepted: 14 Nov 2024.

    Copyright: © 2024 Barbera, Thapa, Adhikari, Li and Lin. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Giovanni Barbera, National Institutes of Health (NIH), Bethesda, United States
    Da-Ting Lin, Behavioral Neuroscience Research Branch, Neural Engineering Section, Intramural Research Program (NIDA), Baltimore, Maryland, United States

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.