Fluorescent recovery after photobleaching (FRAP) was performed on a Zeiss LSM 880 Inverted laser scanning microscope (Zeiss, Oberkochen, Germany) fitted with temperature-controlled and CO2 chamber for live cell work using a Plan-Apochromat 40x/1.3 Oil DIC M27 immersion lens. Images were acquired using ZenBlack operating software. EYFP-YAP expressing cells were seeded in 35 mm glass bottom dishes (MatTek). After 24 h incubation with or without doxycycline, nuclear region was bleached with 405 nm laser wavelength, to reduce nuclear EYFP-YAP signal by at least 70%. Recovery of EYFP-YAP signal inside the nucleus was measured every 5 s over indicated time. Mean fluorescent intensity data was normalized by setting the initial fluorescence signal (pre-bleach) to 100%, and the signal immediately after photo-bleaching (timepoint 0) to 0%. Normalized recovery of EYFP-YAP signal inside the nucleus is presented from timepoint 0s. EYFP-YAP fluorescence in adjacent, non-bleached nuclei remained constant.
Plan apochromat 40x 1.3 oil dic m27 immersion
Manufactured by Zeiss
Sourced in Germany
The Plan-Apochromat 40x/1.3 Oil DIC M27 immersion is a high-quality microscope objective lens manufactured by Zeiss. It features a magnification of 40x and a numerical aperture of 1.3, designed for use with oil immersion microscopy techniques. The lens is part of the Plan-Apochromat series, known for its excellent optical performance and flat field of view.
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2 protocols using plan apochromat 40x 1.3 oil dic m27 immersion
1
Nuclear EYFP-YAP Dynamics Measured by FRAP
2
Nuclear EYFP-YAP Dynamics Measured by FRAP
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Fluorescent recovery after photobleaching (FRAP) was performed on a Zeiss LSM 880 Inverted laser scanning microscope (Zeiss, Oberkochen, Germany) fitted with temperature-controlled and CO2 chamber for live cell work using a Plan-Apochromat 40x/1.3 Oil DIC M27 immersion lens. Images were acquired using ZenBlack operating software. EYFP-YAP expressing cells were seeded in 35 mm glass bottom dishes (MatTek). After 24 h incubation with or without doxycycline, nuclear region was bleached with 405 nm laser wavelength, to reduce nuclear EYFP-YAP signal by at least 70%. Recovery of EYFP-YAP signal inside the nucleus was measured every 5 s over indicated time. Mean fluorescent intensity data was normalized by setting the initial fluorescence signal (pre-bleach) to 100%, and the signal immediately after photo-bleaching (timepoint 0) to 0%. Normalized recovery of EYFP-YAP signal inside the nucleus is presented from timepoint 0s. EYFP-YAP fluorescence in adjacent, non-bleached nuclei remained constant.
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