Development of an automated microscopic system for optogenetics using a mercury lamp fluorescence microscope

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Photomanipulation of genetically encoded photoactivatable proteins, also known as optogenetics, is one of the most innovative techniques in the fields of cell biology. Although photomanipulation is usually performed by diverting the photobleaching mode of a confocal laser microscope, photobleaching by the laser scanning unit is not always suitable for photoactivation. Therefore, we attempted to develop a simple automated wide-field fluorescence microscopy system for optogenetics. An automated fluorescence microscope can be controlled through MetaMorph imaging software, making it possible to acquire time-lapse, multiwavelength images of live cells. We wrote a macro program to change the excitation filter for photoactivation and illumination area during the intervals of image acquisition. When this program was run on the microscope, cells expressing photoactivatable Rac1 showed lamellipodial extension and cell surface ruffling in the illuminated region.