Ix 83 automated inverted microscope

Alexa Fluor® 647 and Alexa Fluor® 568 secondary antibodies were used for dSTORM imaging. The microscope was constructed around an Olympus IX-83 automated inverted microscope. For illumination, an objective-type, total internal reflection fluorescence, oil-immersion objective (APON 60×; NA 1.49 total internal reflection fluorescence; Olympus) was used. A multiline laser source (405, 488, 561, and 640 nm; Andor Technology) was used for excitation and activation. Single-molecular localization signals were separated using appropriate filters (Andor Technology) and detected using electron-multiplying charge-coupled device camera (iXon Ultra 897; Andor Technology). Before dynamic image movie acquisition, conventional fluorescence images were acquired to determine the region of interest. Specifically, 10,000 frame image series were recorded with an exposure time of 50 ms (20 frames per second). The acquired image series were then analyzed using MetaMorph® Super-Resolution System (Molecular Devices) to generate reconstructed dSTORM images.

The microscope was constructed around an Olympus IX-83 automated inverted microscope. For illumination, an objective-type, total internal reflection fluorescence, oil-immersion objective (APON 60×; NA 1.49 total internal reflection fluorescence; Olympus, Tokyo, Japan) was used. A multiline laser source (405, 488, 561, and 640 nm; Andor Technology, Belfast, UK) was used for excitation and activation. Single-molecule localization signals were separated using appropriate filters (Andor Technology) and detected using an electron-multiplying charge-coupled device camera (iXon Ultra 897; Andor Technology). Before dynamic image movie acquisition, conventional fluorescence images were acquired to determine the region of interest. Specifically, 10,000-frame image series were recorded with an exposure time of 50 ms (20 frames per second). The acquired image series were then analyzed using MetaMorph^® Super-Resolution System (Molecular Devices, Silicon Valley, CA, USA) to generate reconstructed STORM images.