hESCs were grown in Matrigel-coated Nunc Lab-Tek chambered coverglass (Thermo Fisher Scientific; cat#155411). Aphidicolin was added at 200 ng/mL for 12 h to synchronize cells in the G1 phase. Before imaging, the cell-permeable Hoechst 33342 dye (Life Technologies) was added to cells at 10–50 ng/mL. Images were captured at 5-min intervals for 10–18 h using a 40X objective and an Evolve 512 Delta EMCCD camera on a Leica inverted microscope equipped with an environmental chamber that controls temperature and CO2. Time-lapsed videos displaying elapsed times between consecutive frames were assembled using the MetaMorph Software (MDS Analytical Technologies) and analyzed using ImageJ. The mitotic duration between nuclear envelope breakdown and anaphase onset was manually determined for each cell. Mitotic defects were visually identified and counted. GraphPad Prism was used to generate the scatter plots and stacked bar graphs.
Evolve 512 delta emccd camera
Manufactured by Leica
The Evolve 512 Delta EMCCD camera is a scientific imaging device designed for low-light applications. It features a high-resolution 512 x 512 pixel sensor and utilizes electron-multiplying CCD (EMCCD) technology to provide enhanced sensitivity and signal-to-noise ratio. The camera is intended for use in various scientific and research environments where the detection and analysis of weak signals are crucial.
Automatically generated - may contain errors
2 protocols using evolve 512 delta emccd camera
1
Monitoring mitotic dynamics in hESCs
2
Monitoring mitotic dynamics in hESCs
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hESCs were grown in Matrigel-coated Nunc Lab-Tek chambered coverglass (Thermo Fisher Scientific; cat#155411). Aphidicolin was added at 200 ng/mL for 12 h to synchronize cells in the G1 phase. Before imaging, the cell-permeable Hoechst 33342 dye (Life Technologies) was added to cells at 10–50 ng/mL. Images were captured at 5-min intervals for 10–18 h using a 40X objective and an Evolve 512 Delta EMCCD camera on a Leica inverted microscope equipped with an environmental chamber that controls temperature and CO2. Time-lapsed videos displaying elapsed times between consecutive frames were assembled using the MetaMorph Software (MDS Analytical Technologies) and analyzed using ImageJ. The mitotic duration between nuclear envelope breakdown and anaphase onset was manually determined for each cell. Mitotic defects were visually identified and counted. GraphPad Prism was used to generate the scatter plots and stacked bar graphs.
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