Na 1.4 plan apo lens

S. pombe strains were grown in YE5S media in the log phase (OD₅₉₅ < 0.8) for 36 h. The cells were harvested at OD₅₉₅ 0.4–0.7 (4–7 × 10⁶ cells/ml), washed twice with EMM5S media, and mounted on EMM5S 2% agarose pads. Images were acquired at room temperature with an Olympus IX-71 microscope with a 100×/NA 1.4 Plan Apo lens (Olympus) and an Andor CSU-X1 spinning disk confocal system with an iXON-EMCCD camera (Andor Technology). Fluorescence was excited using a Coherent OBIS 488-nm LS 20-mW laser with adjustable power and an internal power meter. Time-lapse movies of endocytic actin patches were acquired with Andor iQ2 software (Andor Technology); other images were acquired with the μManager 1.4 plugin (Stuurman et al, 2010 ) for ImageJ (Schneider et al, 2012 (link)).