All cells were imaged using a Nikon Eclipse Ti microscope equipped with Plan Apo 100X (1.45 NA), Plan Apo 60X (1.40 NA), or Plan Fluor 20X (0.5 NA) objectives, an Andor Clara-E camera, and a computer running NIS Elements Software. All cells were viewed in multiple focal planes, and Z-series were captured at 0.2–0.4μm steps. Images presented in the figures represent one slice (Figs 2 ,6 ,7 ) or multiple-slice maximum intensity projections (Figs 3 ,4 ,5 ,8 ).
Plan apo 100x
Manufactured by Oxford Instruments
The Plan Apo 100X is a high-performance microscope objective lens designed for a wide range of applications. It provides a high numerical aperture and flat field of view, delivering exceptional optical performance and resolution. The lens is made with premium optical materials and coatings to ensure optimal image quality and clarity.
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3 protocols using plan apo 100x
1
Fluorescence Imaging of Cellular Structures
2
Microscopic Imaging of Live and Fixed Cells
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All fixed and live cells were imaged using a Nikon Eclipse Ti microscope equipped with Plan Apo 100X (1.45 NA), Plan Apo 60X (1.40 NA), or Plan Fluor 20X (0.5 NA) objectives, an Andor Clara-E camera, and a computer running NIS Elements Software. Most images were taken as single epifluorescence slices, whereas 60X images of mCherry-transfected cells (Fig 10 ) were taken as z-stacks with a 0.3μm step size, and fixed mitotic cells were taken with a 0.3μm (Fig 6 ) or 0.5μm (Figs 5 and 7 ) step size. Live cell imaging was performed in a 35°C chamber (Okolab). During live imaging, cells were cultured in fresh media containing 25mM HEPES (pH 7.4) and DMSO or 4-OHT. Images were captured using the 20x objective at 12min intervals.
3
Live Cell Imaging Protocol for Microscopy
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All fixed and live cells were imaged using a Nikon Eclipse Ti microscope equipped with Plan Apo 100X (1.45 NA), Plan Apo 60X (1.40 NA), or Plan Fluor 20X (0.5 NA) objectives, an Andor Clara-E camera, and a computer running NIS Elements Software. Most images were taken as single epifluorescence slices, whereas 60X images of mCherry-transfected cells (Fig 7 ) were taken as z-stacks with a 0.3µm step size, and fixed mitotic cells were taken with a 0.3µm (Fig 9) During live imaging, cells were cultured in fresh media containing 25mM HEPES (pH 7.4) and DMSO or 4-OHT. Images were captured using the 20x objective at 12min intervals. All image processing and analysis was conducted using ImageJ/FIJI software [100] .
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