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

光操纵基因编码的光活化蛋白质，也称为光遗传学，是细胞生物学领域最具创新性的技术之一。虽然光操纵通常是通过转移的共聚焦激光显微镜的光漂白模式，由激光扫描单元的光漂白并不总是适合于光活化。因此，我们试图开发一种简单的自动化宽视场荧光显微系统的光遗传学。可以通过MetaMorph成像软件控制自动荧光显微镜，从而可以获取活细胞的延时多波长图像。我们编写了一个宏程序，在图像采集的间隔期间改变激发滤光片的光活化和照明区域。当该程序在显微镜上运行时，表达可光活化Rac 1的细胞在照明区域显示板状伪足延伸和细胞表面褶皱。

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.