Apo 20 0.70 dry ph2 objective

Cells were fixed with 0.5% glutaraldehyde at room temperature for 15 min. After that, cells were washed twice with 1 mM MgCl₂/PBS and incubated with X-Gal staining solution at 37° C for 3 hours. The staining was terminated by three consecutive washes with ddH₂O. Finally, the cells were let dry, mounted with Mowiol containing DAPI and imaged on the Leica DM6000 fluorescent microscope using the HC PLAN APO 20×/0.70 DRY PH2 objective and color CCD camera Leica DFC490 (Leica Microsystems GmbH, Wetzlar, Germany).

The development of cellular senescence was followed by the determination of senescence-associated beta-galactosidase activity (Dimri et al., 1995 (link)). Cells were fixed with 0.5% glutaraldehyde at room temperature for 15 min. Cells were then washed twice with 1 mM MgCl₂/PBS and incubated with X-Gal staining solution at 37°C for 3 h. The staining was terminated by three consecutive washes with ddH₂O. Finally, the cells were let dry, mounted with Mowiol containing DAPI, and imaged on the Leica DM6000 fluorescent microscope using the HC PLAN APO 20×/0.70 DRY PH2 objective and color CCD camera Leica DFC490 (Leica Microsystems GmbH, Wetzlar, Germany).

Cells were fixed with 0.5% glutaraldehyde at RT for 15 min at indicated time-points. After that, cells were washed twice with 1 mM MgCl₂/PBS and incubated with X-Gal staining solution for 3 hours at 37°C. The staining was terminated by three consecutive washes with ddH₂O. Finally, the cells were let dry, mounted with Antifade Pro-long Gold Mounting Media and imaged on the Leica DM6000 fluorescent microscope using the HC PLAN APO 20×/0.70 DRY PH2 objective and color CCD camera Leica DFC490 (Leica Microsystems GmbH, Wetzlar, Germany).