As previously detailed [28 (link), 31 (link)], confocal images were acquired in a FV1000 laser scanning microscope (Olympus) using an Olympus UPlanSApo 60x oil immersion objective (NA = 1.35). Cy3 were excited using a He-Ne green laser at 543 nm (average power at the sample, 700 nW). Fluorescence was detected with a photomultiplier set in the pseudo photon-counting detection mode, using 560–660 nm filtering for Cy3 detection. An averaged image was obtained from 10 consecutive images acquired per cell.
Uplansapo 60x oil
Manufactured by Olympus
The UPlanSApo 60x oil is an objective lens designed for use in optical microscopy. It features a high numerical aperture and is optimized for use with oil immersion, providing high-resolution imaging capabilities.
Automatically generated - may contain errors
2 protocols using uplansapo 60x oil
1
Confocal Imaging of Cy3-Labeled Cells
2
Imaging of Extracellular Vesicles in NSCs
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Following addition of labeled EVs or filtered culture media, cells were briefly fixed (2 % PFA, 15 min). NSC nuclei were counterstained with 300 nM DAPI (Thermo Fisher; Catalog #D1306), mounted onto glass slides (Vectashield, Vector Laboratories; Catalog # H-1200-10; Burlingame, CA/USA), coverslipped, and imaged using a confocal-laser scanning microscope (FluovView-1200, Olympus Corporation of the Americas; Center Valley, PA/USA) equipped with a 405 nm laser to excite DAPI and a 559 nm laser to excite MemBrite 568/580. Micrographs were acquired using a 60x magnification objective (UPlanSApo 60X Oil, Olympus) with 2x zoom through image spatial resolution adjustment.
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