optoPlate-96

The optoPlates are a series of open-source devices for high-throughput, multi-color optogenetic experiments in microwell plates. Each well of a 96-well plate can be illuminated with up to 3 wavelengths of light, with independent control of illumination timing and intensity. Despite its complexity and robustness, the optoPlate-96 can be assembled in a few hours with no specialized equipment, at a total cost of ~$500. See below for full optoPlate-96 details, annotated protocols, design files, and code. These devices have been game-changers in our ability to perform complex, quantitative, and reproducible optogenetic experiments. We hope they will be equally powerful for you.

photo credit: ©Steve Babuljak


Key References

Bugaj LJ and Lim WA. (2019). High throughput multicolor optogenetics in microwell plates. Nature Protocols.
**please cite this work if you use the optoPlates in a publication

Bugaj, L.J., Sabnis, A.J., Mitchell, A., Garbarino, J.E., Toettcher, J.E., Bivona, T.G., and Lim, W.A. (2018). Cancer mutations and targeted drugs can disrupt dynamic signal encoding by the Ras-Erk pathway. Science 361.


IMG_8594.jpg

Overview

At its core, the optoPlate-96 is an LED array formatted to illuminate standard microwell plates. This device allows programmable control of 288 independent channels of light in 96-well format (3 channels/colors per well). Each well position can also illuminate 4-wells of a 384-well plate, and the devices can be readily multiplexed for enhanced throughput. 3D-printed adapters were designed to optically insulate each well from its neighbors. Additionally, an active heatsink minimizes sample heating when light-intensive illumination protocols must be used. The optoPlates were designed primarily for use within cell culture incubators, but can also be adapted for illumination of cells during live-cell microscopy.

IMG_8597.jpg

optoPlate-96 features

  • 96 illumination positions

  • up to 3 colors per condition. optimized wavelengths for common blue and red mammalian optogenetic probes (blue/red/far-red illumination)

  • independent control of each color/position

  • 96- or 384-well plate illumination

  • 3D-printed microwell plate adapter and lid

  • complete optical insulation in each well

  • designed for long-term incubator experiments

  • thermal management

  • fully integrated, no data cables

  • powered by Arduino

  • simple DIY assembly, QC in 3-4 hours

IMG_8598.jpg

Assembly and use

The optoPlate-96 can be built to illuminate 1, 2 or 3 separate colors per position. Our protocol provides instructions on how to build any of three color combinations that can stimulate the common mammalian cell optogenetic probes:

  1. Blue (for LOV2, Cryptochrome2)

  2. Red/Far-red (for Phytochrome B)

  3. Blue/Red/Far-red (for multiplexing 1 and 2)

The optoPlate-96 can be assembled manually following our detailed protocol.  Once all necessary components have been ordered/3D printed/laser cut, a fully functional optoPlate-96 can be assembled and tested in 3-4 hours. The design files for PCB printing, 3D-printing, and laser/jet cutting are all provided in our Github repository.  To visualize the finished product, explore the interactive 3D models below.


Downloads


Interactive 3D models

click on an image to interact with the model


Comments? Questions? Bugs?

We would love feedback on the optoPlate-96. What works? What doesn’t? What could be done better?. Drop us a note below and we will get back to you as soon as we can.

Name *
Name