Cells were incubated with 10 μM 5-ethynyl-2’-deoxyuridine (EdU) for 24 hours and fixed with 4% formaldehyde at room temperature for 15 min. To visualize EdU incorporation, click chemistry was performed with Click-iT™ EdU Cell Proliferation Kit with Alexa Fluor™ 647 dye (Thermo Fisher Scientific, Waltham, MA, USA) according to manufacturer’s instructions. The stained cells were acquired by high-content imaging using inverted wide-field microscope (Olympus IX81) equipped with UPLXAPO 20×/0.8 DRY CORR; FWD 0.6; CG 0 – 2 objective and sCMOS camera Hamamatsu ORCA-Flash4.0 LT+90, 6.5 μm pixel.
Orca flash4.0 lt 90
Manufactured by Hamamatsu Photonics
The ORCA-Flash4.0 LT+90 is a scientific-grade digital CMOS camera designed for low-light imaging applications. It features a large 4.2-megapixel sensor, a fast readout speed of up to 100 frames per second, and a high quantum efficiency for improved light sensitivity. The camera is equipped with a Peltier cooling system to reduce thermal noise and support long exposure times. It is compatible with a variety of scientific imaging techniques and is suitable for a wide range of applications in fields such as microscopy, spectroscopy, and bioluminescence imaging.
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2 protocols using orca flash4.0 lt 90
1
Visualizing Cell Proliferation with EdU Staining
2
Quantifying Cell Proliferation via EdU Labeling
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Cells were incubated with 10 μM 5-ethynyl-2’-deoxyuridine (EdU) for 24 h and fixed with 4% formaldehyde at room temperature for 15 min. To visualize EdU incorporation, click chemistry was performed with Click-iT EdU Cell Proliferation Kit with Alexa Fluor™ 647 dye (Thermo Fisher Scientific, Waltham, MA, USA) according to the manufacturer’s instructions. The stained cells were acquired by high-content imaging using an inverted wide-field microscope (Olympus IX81) equipped with UPLXAPO 20×/0.8 DRY CORR; FWD 0.6; CG 0–2 objective and sCMOS camera Hamamatsu ORCA-Flash4.0 LT + 90, 6.5 μm pixel.
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