Grasshopper3 cmos camera

Transfected HEK293 cells were examined with an SP8 confocal fluorescence microscope using 63 × 1.4 NA objective, Leica (Wetzlar, Germany). Alternatively, cells were observed microscopically using an upright Olympus BX51WI equipped with 100 × 1 NA objective and images were captured with a Grasshopper3 CMOS camera (FLIR, Richmond, BC, Canada) controlled by manufacturer provided software. To generate images for quantitative analysis, we kept camera, microscope, and excitation light source (X-Cite 120LEDBoost, Excelitas Technologies, 2% intensity) settings constant. To observe cells with low-level fluorescence, excitation light was set at 15% of intensity. Fluorescence intensities were obtained using Fiji^{37 (link)}. Regions of interest were used to obtain average intensity values from the transfected HEK293 cells (I_cell) and an area without cells (I_background) within the same image. Fluorescence intensity of transfected HEK293 cells (I_norm) for each cell was normalized (I_norm = I_cell − I_background). Each data sample represent average normalized intensity of the transfected cells within an image. We captured 3 to 6 fields of view per dish, using two dishes per construct in every experiment.

Transfected HEK293 cells were examined with an SP8 confocal fluorescence microscope using 63 × 1.4 NA objective, Leica (Wetzlar, Germany) as described previously^{17 (link)}. Briefly, cells were observed microscopically using an upright Olympus BX51WI equipped with 100 × 1 NA objective and images were captured using a Grasshopper3 CMOS camera (FLIR, Richmond, BC, Canada) controlled by Leica LAS X version 3.5 software (available at: https://www.leica-microsystems.com/products/microscope-software/p/leica-las-x-ls/). All figures were created using Adobe Illustrator CS6 (Available at: https://adobe.com/products/illustrator), under an Adobe Inc., Creative Cloud Desktop 2019 shared device license to Case Western Reserve University (CWRU) that operates until 3/31/2022.