# Enhancing high-throughput optogenetics: Integration of LITOS with Lustro enables simultaneous light stimulation and shaking

## TL;DR

This paper describes a new setup combining LITOS and Lustro to allow simultaneous light stimulation and shaking in optogenetic experiments.

## Contribution

A 3D-printed adaptor enables LITOS compatibility with the BioShake 3000-T ELM for concurrent illumination and shaking.

## Key findings

- Constant shaking increased the maturation rate of the mScarlet-I fluorescent reporter in yeast cultures.
- Modified LITOS with the adaptor caused only a marginal temperature increase during operation.
- The new setup is compatible with automated platforms like Lustro for high-throughput optogenetic experiments.

## Abstract

Optogenetics is a powerful tool that uses light to control cellular behavior. Here we enhance high-throughput characterization of optogenetic experiments through the integration of the LED Illumination Tool for Optogenetic Stimulation (LITOS) with the previously published automated platform Lustro. Lustro enables efficient high-throughput screening and characterization of optogenetic systems. The initial iteration of Lustro used the optoPlate illumination device for light induction, with the robot periodically moving the plate over to a shaking device to resuspend cell cultures. Here, we designed a 3D-printed adaptor, rendering LITOS compatible with the BioShake 3000-T ELM used in Lustro. This novel setup allows for concurrent light stimulation and culture agitation, streamlining experiments. Our study demonstrates comparable growth rates between constant and intermittent shaking of
Saccharomyces cerevisiae
liquid cultures. While the light intensity of the LITOS is not as bright as the optoPlate used in the previous iteration of Lustro, the constant shaking increased the maturation rate of the mScarlet-I fluorescent reporter used. Only a marginal increase in temperature was observed when using the modified LITOS equipped with the 3D-printed adaptor. Our findings show that the integration of LITOS onto a plate shaker allows for constant culture shaking and illumination compatible with laboratory automation platforms, such as Lustro.