RRC ID 67879
著者 Araki N, Ikeda Y, Kato T, Kawai K, Egami Y, Miyake K, Tsurumaki N, Yamaguchi M.
タイトル Development of an automated fluorescence microscopy system for photomanipulation of genetically encoded photoactivatable proteins (optogenetics) in live cells.
ジャーナル Microscopy (Oxf)
Abstract Photomanipulation of genetically encoded light-sensitive protein activity, also known as optogenetics, is one of the most innovative recent microscopy techniques in the fields of cell biology and neurobiology. 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. We have developed a simple automated wide-field fluorescence microscopy system for the photomanipulation of genetically encoded photoactivatable proteins in live cells. An electrically automated fluorescence microscope can be controlled through MetaMorph imaging software, making it possible to acquire time-lapse, multiwavelength images of live cells. Using the journal (macro recording) function of MetaMorph, 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 wide-field fluorescence microscope, cells expressing genetically encoded photoactivatable Rac1, which is activated under blue light, showed morphological changes such as lamellipodial extension and cell surface ruffling in the illuminated region. Using software-based development, we successfully constructed a fully automated photoactivation microscopy system for a mercury lamp-based fluorescence microscope.
巻・号 63(3)
ページ 255-60
公開日 2014-6-1
DOI 10.1093/jmicro/dfu003
PII dfu003
PMID 24523516
MeSH Animals Automation, Laboratory / methods Cells / ultrastructure* Mice Microscopy, Fluorescence / methods* Optogenetics / methods* rac1 GTP-Binding Protein / genetics rac1 GTP-Binding Protein / ultrastructure
IF 1.394
リソース情報
ヒト・動物細胞 RAW 264(RCB0535)