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  1. Home
  2. Voltage Imaging With Periodic Structured Illumination.
  1. Home
  2. Voltage Imaging With Periodic Structured Illumination.

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Voltage imaging with periodic structured illumination.

Forest Speed1, Alec Teel2, Timothy Young1

  • 1Department of Bioengineering, University of Colorado Anschutz Medical Campus, Aurora, CO 80045, USA.

Biomedical Optics Express
|January 14, 2026

View abstract on PubMed

Summary
This summary is machine-generated.

Periodic structured illumination with pseudo-HiLo (pHiLo) image reconstruction reduces out-of-focus signals in voltage imaging. This method improves signal clarity for in-focus cells compared to traditional widefield recordings.

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Area of Science:

  • Neuroscience
  • Optical Imaging
  • Biophysics

Background:

  • In vivo voltage imaging is crucial for understanding neural circuit dynamics.
  • Out-of-focus cellular signals can contaminate recordings, reducing the accuracy of voltage activity measurements.
  • Traditional widefield imaging and pseudo-widefield (pWF) reconstructions struggle to mitigate this out-of-focus light effectively.

Purpose of the Study:

  • To evaluate the efficacy of periodic structured illumination with pseudo-HiLo (pHiLo) image reconstruction for in vivo voltage imaging.
  • To compare the performance of pHiLo against traditional widefield recordings and pseudo-widefield (pWF) reconstructions in reducing out-of-focus signals.
  • To assess the impact of pHiLo on key imaging metrics such as spike peak-to-noise ratio (PNR) and cell-background correlation.

Main Methods:

  • Utilized periodic structured illumination with pseudo-HiLo (pHiLo) image reconstruction.
  • Performed in vivo voltage imaging experiments.
  • Compared pHiLo with traditional widefield illumination and pseudo-widefield (pWF) reconstructions.
  • Analyzed spike peak-to-noise ratio (PNR) and cell-background correlation for time courses.

Main Results:

  • pHiLo significantly reduced signal from out-of-focus cells compared to traditional widefield recordings and pWF reconstructions.
  • Demonstrated improved clarity of voltage activity for in-focus cells of interest using pHiLo.
  • Quantitative comparisons of PNR and cell-background correlation were made between pHiLo and soma-targeted widefield illumination (TI).

Conclusions:

  • pHiLo is an effective technique for reducing out-of-focus light contamination in high-speed in vivo voltage imaging.
  • The study highlights the tradeoffs between out-of-focus light reduction, signal-to-noise ratio, and temporal resolution with pHiLo.
  • pHiLo offers advantages for imaging neural activity in awake mice by enhancing signal quality.