Pln apo100x 1.46 oil

Samples were examined in a Zeiss Axioimager Z.1 platform equipped with the Elyra PS.1 super-resolution module for structured illumination (SIM) and the LSM780 module for CLSM. High resolution images were acquired in super-resolution mode using Zeiss Pln Apo100x/1.46 oil objective (tot. mag. 1600x) with appropriate oil (Immersol 518F). SR-SIM setup involved 5 rotations and 5 phases for each image layer and up to 7 Z-stacks (101nm) were acquired per image. CLSM setup for FRAP and life cells acquisition involved c-Apo 40x/1.2W water immersion objective. Bleaching of regions of interest (ROI) was performed using Argon 488nm laser. Lower resolution images of fixed samples were acquired using Plan Apo 63x/1.4 Oil objective (tot. mag. 1008x). FRAP and image acquisitions were performed in Zeiss Zen 11 software. For FRAP internal Zen’s “Bleach” and “Regions” modules were used. Data from FRAP analysis involving multiple bleached ROI’s were exported into MS-Excel and charted. Basic processing of acquired images such as contrast and brightens setting was done in Adobe Photoshop on images exported as tiff. Quantitative microscopy-based cytometry of the IF stained samples was performed using an automatic inverted fluorescence microscope BX71 (Olympus) in the ScanR Acquisition software (Olympus), analyzed with ScanR Analysis software (Olympus).