Engineering LED photoconversion chambers
This project is aimed at constructing LED-based photoconversion chambers to photoactivate/convert fluorescent and optogenetic proteins and biosensors in cultured cells and ex vivo brain tissue. The ideal system would contain modular LED arrays to deliver homogenous fixed wavelength illumination to cultured cells and ex vivo tissue preparations with the purpose of exciting/activating/converting light-based protein tools for screening and evolution of new biosensor systems and optogenetic proteins. The chamber would be constructed in a manner that incorporates LED arrays with mirrored surfaces for homogenous light-dispersion, be modular with the ability to interchange LED arrays across multiple different wavelengths, be compact for housing in cell/tissue culture incubators for environmental control, and interface with programmable software to write stimulation protocols controlling LED output power, and illumination duration and frequency. This type of photoconversion chamber would greatly accelerate screening applications associated with large libraries of newly engineered or evolved protein-based fluorescent tools and overcome the bottleneck of conventional microscopy-based illumination which typically restricts illumination area and is not suited for batch conversion needs. My laboratory, in particular, is constructing new optogenetic proteins and biosensors to sense and manipulate glial signaling cascades which will substantially advance knowledge surrounding neuro-glial communication in the healthy and diseased brain. However, we currently are restricted in screening large protein libraries that prevents high-throughput engineering and directed evolution of these new cutting-edge tools.