HeLa LAP2b:RFP or tubulin:GFP cells were seeded onto Petri dishes with a 30-mm glass base (Greiner), and an inverted fluorescence microscope (Observer D1; Zeiss) was used. Fluorescence or phase-contrast images were taken by using a 63× Plan Neofluor 0.75 NA Ph2 dry objective (Zeiss). Imaging was performed at 37°C in 5% CO2 by using a Zeiss AxioCam MRm camera and Zeiss ZEN 2 acquisition software. Drugs were added to the medium immediately before filming as appropriate.
30 mm glass base
Manufactured by Greiner
The 30-mm glass base is a laboratory equipment item used as a foundation or support for various experimental setups. It provides a stable and level surface for placing and securing other laboratory equipment or apparatus. The glass base measures 30 millimeters in diameter and is designed to withstand the weight and demands of laboratory work.
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4 protocols using 30 mm glass base
1
Fluorescence Microscopy of Live Cells
2
Live-Cell Fluorescence Microscopy
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HeLa or BE cells were seeded onto Petri dishes with a 30-mm glass base (Greiner). An inverted fluorescence microscope (Observer D1; Zeiss) was used. Fluorescence images were taken by using a 63× Plan neofluar 0.75 NA Ph2 objective (Zeiss) and phase-contrast images with a 20× Plan neofluar 0.40 NA Ph2 objective (Zeiss). Imaging was performed in air, at 37°C in 5% CO2, by using a Zeiss AxioCam MRm camera and the Zeiss ZEN2 acquisition software.
3
Imaging live cells using fluorescence microscopy
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BE or HeLa LAP2b-RFP cells were seeded onto Petri dishes with a 30-mm glass base (Greiner), and an inverted fluorescence microscope (Observer D1; Zeiss) was used. Fluorescence and phase-contrast images were taken by using a 63× Plan Neofluor 0.75 NA Ph2 dry objective (Zeiss; Figs. 1 E and S1 C), a 20× Plan Neofluor 0.40 NA Ph2 dry objective (Zeiss; Fig. 7, C and D ), or a 10× A-plan 0.25 Ph1 dry objective (Zeiss; Fig. 7, E and F ). Imaging was performed at 37°C in 5% CO2 by using a Zeiss AxioCam MRm camera and the Zeiss ZEN 2 acquisition software. For UCN-01 or PP2 treatments in Figs. 1 and 7 , the drugs were added to the medium immediately before filming.
4
Fluorescent Imaging of Live Cell Dynamics
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HeLa LAP2b:RFP or HeLa tubulin:GFP cells were seeded onto Petri dishes with a 30-mm glass base (Greiner) and an inverted fluorescence microscope (Observer D1; Zeiss) was used. Fluorescence or phase-contrast images were taken by using a 63× Plan Neofluor 0.75 NA Ph2 dry objective (Zeiss). Imaging was performed at 37°C in 5% CO2 by using a Zeiss AxioCam MRm camera and Zeiss ZEN 2 acquisition software. Drugs were added to the medium immediately before filming as appropriate.
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