High-throughput feedback-enabled optogenetic stimulation and spectroscopy in microwell plates.

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.