Automated Phototransfection

This project is on automating the phototransfection process on neuron and fibroblast cells. Phototransfection is presently done manually in a very tedious manner: First, the cell of interest needs to be identified in the microscope FOV and location recorded. Then an appropriate location of where to administer a laser beam on the cell body needs to be determined. Applying the laser to the wrong location can damage the cell. The laser beam creates a hole in the cell membrane where donor mRNA can be inserted to the cell from a pipette, completing the process. A framework for fully automating this procedure has been designed and proof-of-concept implementation achieved. Computer vision techniques are used to identify the cell of interest in the FOV and determine target locations for the laser beam. A control program takes this information and coordinates movements of the computer controlled XY stage, translating the coordinates of the laser target location to a predefined, fixed, laser firing location. A 20X improvement was shown to be possible with this implementation with room for improvement to greater than 80X.