The experimental approach for immunofluorescence microscopy are adapted from previous literature (64 (link)). Briefly, HEK293 cells were cultured on poly-lysine coated coverslips, transfected with plasmids expressing FLAG-tagged HO2 and its variants, washed with phosphate buffered saline (PBS), and fixed with 4% w/v paraformaldehyde before immunostaining. Immunofluorescence microscopy samples were permeabilized with 1× PBS containing 0.2% v/v Triton X-100 and stained with corresponding antibodies. Sample images were acquired on an Olympus IX81 microscope with a Yokogawa CSU-W1 spinning disk confocal scanner unit, an Olympus PlanApo 60× 1.42 NA objective, and a Hamamatsu ImagEMX2-1K EM-CCD digital camera.
Imagemx2 1k em ccd digital camera
Manufactured by Hamamatsu Photonics
The ImagEMX2-1K is a high-performance EM-CCD digital camera manufactured by Hamamatsu Photonics. It features a 1024 x 1024 pixel sensor with a pixel size of 13 x 13 μm. The camera offers a maximum readout rate of 17 MHz and a peak quantum efficiency of over 95%. It is capable of producing high-quality, low-noise images, making it suitable for a variety of scientific and industrial applications.
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2 protocols using imagemx2 1k em ccd digital camera
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Immunofluorescence Imaging of HO2 Variants
2
Immunofluorescence protocol for HEK293 cells
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The experimental approach for IF are adapted from previous literature (64) . Briefly, HEK293 cells were cultured on poly-lysine coated coverslips, transfected with plasmids expressing FLAG-tagged HO2 and its variants, washed with phosphate buffered saline (PBS) and fixed with 4% paraformaldehyde (w/v) before immunostaining. IF samples were permeabilized with 1x PBS containing 0.2% Triton X-100 (v/v) and stained with corresponding antibodies. Sample images were acquired on an Olympus IX81 microscope with a Yokogawa CSU-W1 spinning disk confocal scanner unit, an Olympus PlanApo 60x 1.42 NA objective, and a Hamamatsu ImagEMX2-1K EM-CCD digital camera.
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