Miniature structured illumination microscope for in vivo 3D imaging of brain structures with optical sectioning

Omkar D Supekar; Andrew Sias; Sean R Hansen; Gabriel Martinez; Graham C Peet; Xiaoyu Peng; Victor M Bright; Ethan G Hughes; Diego Restrepo; Douglas P Shepherd; Cristin G Welle; Juliet T Gopinath; Emily A Gibson

doi:10.1364/BOE.449533

Miniature structured illumination microscope for in vivo 3D imaging of brain structures with optical sectioning

Biomed Opt Express. 2022 Mar 29;13(4):2530-2541. doi: 10.1364/BOE.449533. eCollection 2022 Apr 1.

Authors

Omkar D Supekar^{1

2

3}, Andrew Sias⁴, Sean R Hansen⁴, Gabriel Martinez⁴, Graham C Peet⁵, Xiaoyu Peng⁶, Victor M Bright², Ethan G Hughes⁵, Diego Restrepo⁵, Douglas P Shepherd⁷, Cristin G Welle^{6

8}, Juliet T Gopinath^{1

9}, Emily A Gibson^{4

10}

Affiliations

¹ Department of Electrical, Energy and Computer Engineering, University of Colorado Boulder, CO 80309, USA.
² Department of Mechanical Engineering, University of Colorado Boulder, CO 80309, USA.
³ omkar.supekar@colorado.edu.
⁴ Department of Bioengineering, University of Colorado Anschutz Medical Campus, CO 80045, USA.
⁵ Department of Cell and Developmental Biology, University of Colorado Anschutz Medical Campus, CO 80045, USA.
⁶ Department of Physiology and Biophysics, University of Colorado Anschutz Medical Campus, CO 80045, USA.
⁷ Department of Physics, Arizona State University, Tempe, AZ 85287, USA.
⁸ Department of Neurosurgery, University of Colorado Anschutz Medical Campus, CO 80045, USA.
⁹ Department of Physics, University of Colorado Boulder, CO 80309, USA.
¹⁰ emily.gibson@cuanschutz.edu.

Abstract

We present a high-resolution miniature, light-weight fluorescence microscope with electrowetting lens and onboard CMOS for high resolution volumetric imaging and structured illumination for rejection of out-of-focus and scattered light. The miniature microscope (SIMscope3D) delivers structured light using a coherent fiber bundle to obtain optical sectioning with an axial resolution of 18 µm. Volumetric imaging of eGFP labeled cells in fixed mouse brain tissue at depths up to 260 µm is demonstrated. The functionality of SIMscope3D to provide background free 3D imaging is shown by recording time series of microglia dynamics in awake mice at depths up to 120 µm in the brain.

Associated data

figshare/10.6084/m9.figshare.19142336

Abstract

Associated data

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