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An Optogenetic Method to Control and Analyze Gene Expression Patterns in Cell-to-cell Interactions
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High-throughput feedback-enabled optogenetic stimulation and spectroscopy in microwell plates.

William Benman1, Saachi Datta1, David Gonzalez-Martinez1

  • 1Department of Bioengineering, University of Pennsylvania, Philadelphia, PA, 19104, USA.

Communications Biology
|November 24, 2023
PubMed
Summary

Researchers developed the optoPlateReader (oPR), an open-source device for real-time monitoring of optogenetic experiments. This innovation enables automated stimulation, spectroscopy, and feedback control in microwell plates, advancing high-throughput biological research.

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Area of Science:

  • Biotechnology
  • Optogenetics
  • Microfluidics

Background:

  • Sophisticated optogenetic experiments are limited by the lack of real-time monitoring tools.
  • Current methods require sample transfers to separate instruments, hindering high-throughput analysis.
  • Open-source illumination devices offer programmable light patterns but lack integrated measurement capabilities.

Purpose of the Study:

  • To develop an open-source device for automated optogenetic stimulation and real-time monitoring in microwell plates.
  • To integrate illumination and spectroscopic measurement capabilities into a single, compact instrument.
  • To enable computer-in-the-loop feedback control for optogenetic experiments.

Main Methods:

  • Development of the optoPlateReader (oPR), integrating an illumination module and a 96-channel optoReader for parallel measurements.
  • Optimization for blue light stimulation and optical density/fluorescence measurements.
  • Calibration of all device components for accurate data acquisition.

Main Results:

  • Demonstrated oPR's capability for real-time growth measurement and fluorescent protein expression induction in E. coli.
  • Successfully implemented computer-in-the-loop feedback control by adjusting stimulation based on sample state.
  • Validated the device's performance for automated optical read/write functions in 96-well plates.

Conclusions:

  • The optoPlateReader (oPR) addresses the need for integrated, real-time monitoring in optogenetic microwell plate experiments.
  • Open-source, accessible hardware enables automated optical read/write and feedback control, expanding experimental possibilities.
  • The oPR facilitates advanced, high-throughput optogenetic research through customizable and cost-effective instrumentation.