the optoPlate

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 for each wavelength. We designed the optoPlates for robust operation in standard cell culture incubators, and for straightforward assembly and use. The optoPlate-96 can be assembled in a few hours with no specialized equipment, at a total cost of ~$600. See below for full device details, annotated protocols, design files, and code. These devices have enabled us to perform complex, quantitative, and reproducible optogenetic experiments over timescales ranging from minutes to weeks. We hope you will find them equally enabling.

(photo credit: ©Steve Babuljak)

OVERVIEW

the optoPlate

An optoPlate is, in essence, 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.

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

  • DIY assembly, QC in 3-4 hours

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 (LOV2, Cryptochrome2, etc.)

  2. Red/Far-red (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

ORDER

The optoPlate is designed for rapid assembly and use by non-specialists (see Downloads and User Additions). However, if you would rather purchase an optoPlate, we have partnered with LABmaker, a company that assembles and distributes DIY open-science hardware.

INTERACTIVE 3D MODELS

click on an image to interact with the model

OptoPlate users have created independent software and protocols to further empower these devices (thank you!!). Check out their work in the links below. And if you’ve made further modifications in software, hardware, or process, and would like to share with the optoPlate community, please get in touch and we’ll be happy to feature your work here.

Supplementary Assembly Manual. Mary Dunlop has put together a companion assembly protocol complete with videos, tips, and more detailed guidance on how to perform some of the more complex assembly steps. Highly recommended to read this before your first assembly — thank you Mary! Mary also filmed this beautiful assembly video:

USER ADDITIONS

credit: Mary Dunlop, PhD, Boston University

Graphical user interface (GUI). The Hoerner and Weber group have developed an intuitive GUI to allow easy point-and-click programming of complex 3-color illumination. Check out their protocol and software.

Automated calibration of LED intensities. Due to variability in manufacturing and assembly, light output from individual LEDs can vary by ~10-20%. Thus to achieve consistent grey-scale intensity modulation between wells, all LEDs on an optoPlate illuminator should be calibrated. Luckily, the McClean group has automated this otherwise tedious calibration process (protocol and software).

Please cite the following work if you use the optoPlates in a publication

Bugaj LJ and Lim WA. (2019). High throughput multicolor optogenetics in microwell plates. Nature Protocols. doi:10.1038/s41596-019-0178-y link

To see a list of papers that have used and/or cited the optoPlate, click here (Google Scholar)

CITATION

COMMENTS? QUESTIONS? BUGS?

We would love your feedback on the optoPlate. Drop us a note below and we will get back to you as soon as we can.