Dfc365 fx digital camera

Fluorescence microscopy was performed on a Leica DM6000B Microscope, equipped with a Leica DFC365 FX digital camera. Images were captured using the Leica Application Suite (LAS-AF) microscope software. Representative pictures were taken using a 40x/0,85 Leica HC PL-APOCHROMAT objective.
For quantifications of INL thickness and TUNEL assays, pictures were captured in the posterior pole of the retina, once ventrally and once dorsally to the optic nerve head. Three sagittal sections from three and four different animals for each genotype and age were analysed for INL thickness measurement and TUNEL assay respectively.

Following exposure to PL, cells were stained with 10 μM of the dye dihydrodichlorofluorescein-diacetate (H2DCFDA, Invitrogen Inc., Eugene, OR, USA) in 1 ml media to measure intracellular hydrogen peroxide. Cells were stained for 30 min at 37°C.
Fluorescence microscopy was performed on a Leica DM6000B Microscope, equipped with a Leica DFC365 FX digital camera. Images were captured using the Leica Application Suite (LAS-AF) microscope software. Representative pictures were taken using a 40x/0,85 Leica HC PL-APOCHROMAT objective.

Overnight cultures of individual strains were spun down (5,000 × g, 5 min), resuspended to an OD₆₀₀ of 1.0 in PBS-T, and transferred in 500 µl aliquots into 2 ml microcentrifuge tubes. 50 µg of GFP-tagged RBP was added to the cells and incubated for 90 min on an overhead rotator. Cells were spun down and washed once with 1 ml PBS-T and resuspended in 200 µl PBS-T. 150 µl of the cell suspension was pipetted into a well of a black, flat bottom 96-well microplate (Greiner Bio-One, Austria) and the fluorescence intensity of bound GFP-RBP was measured at ambient temperature using a POLARStar Omega spectrophotometer (BMG Labtech, Germany) at 485 nm excitation, 520 nm emission with (1000 x) fixed gain. Fluorescence binding was performed in triplicate with mean (raw fluorescence) ± standard deviation. For fluorescence microscopy, 4 μl of the cell suspension was imaged using a confocal inverted microscope (Leica TCS SPE) equipped with an ACS APO 63×/1.30 oil CS lens objective with excitation at 488 nm and emissions collected with a PMT detector in the detection range of 510 to 550 nm. Transmitted-light microscopy images were obtained with the differential interference contrast mode. Images were acquired with a Leica DFC 365 FX digital camera controlled with the LAS AF software. Fiji v2.0.0 (ImageJ software) was used to generate final images.