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Updated: Sep 12, 2025

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
Published on: May 30, 2016
Voltage Imaging with Periodic Structured Illumination
Forest Speed1, Alec Teel2, Gregory L Futia1
1Department of Bioengineering, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.
Abstract:
We utilize periodic structured illumination with pseudo-HiLo (pHiLo) image reconstruction for in vivo voltage imaging. We demonstrate reduced signal from out-of-focus cells, that contaminates voltage activity for in-focus cells of interest, with pHiLo compared to traditional widefield recordings taken with uniform illumination and pseudo-widefield (pWF) reconstructions. We discuss tradeoffs between signal-to-background ratio, signal-to-noise ratio and temporal resolution for pHiLo in the context of high-speed voltage imaging in awake mice.

