Three-dimensional multiphoton imaging of brain activity in freely-moving mice using a miniature microscope with variable focus lens (Conference Presentation)

Emily A. Gibson; Baris N Ozbay; Gregory L Futia; Ming Ma; Ethan G Hughes; Diego Restrepo

doi:10.1117/12.2309388

14 March 2018 Three-dimensional multiphoton imaging of brain activity in freely-moving mice using a miniature microscope with variable focus lens (Conference Presentation)

Emily A. Gibson, Baris N Ozbay, Gregory L Futia, Ming Ma, Ethan G Hughes, Diego Restrepo

Author Affiliations +

Proceedings Volume 10499, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV; 104991K (2018) https://doi.org/10.1117/12.2309388
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

We report a miniature head mounted two-photon fiber-coupled microscope (TP-FCM) for neuronal imaging with active axial focusing enabled using a miniature electrowetting lens. Full three-dimensional two-photon imaging of GCaMP6s showing individual neuron activity in multiple focal planes was achieved in a freely-moving mouse. Two-color simultaneous imaging of GFP and tdTomato fluorescence is demonstrated. Additionally, the axial scanning of the electrowetting lens allows dynamic control of tilt to the focal plane allowing rapid scanning of different regions of interest in three dimensions. Two-photon imaging allows increased penetration depth in tissue with a field-of-view of 240 μm diameter and 200 μm variable axial focus. The TP-FCM has a light-weight design (~4 g) and excellent image stability. TP-FCM with dynamic axial scanning provides a new capability to record from functionally distinct neuronal layers, opening up unique opportunities in neuroscience research.

Conference Presentation

Citation Download Citation

Emily A. Gibson, Baris N Ozbay, Gregory L Futia, Ming Ma, Ethan G Hughes, and Diego Restrepo "Three-dimensional multiphoton imaging of brain activity in freely-moving mice using a miniature microscope with variable focus lens (Conference Presentation)", Proc. SPIE 10499, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXV, 104991K (14 March 2018); https://doi.org/10.1117/12.2309388

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KEYWORDS

3D image processing

Microscopes

Brain

Neuroimaging

Two photon imaging

Green fluorescent protein

Head

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