Two-photon imaging microscopy was performed to obtain 2D representations of the two Saccharomyces cerevisiae strains. The images were collected on a Nikon A1-MP scanning microscope equipped with a Plan APO IR × 60 objective (NA, 1.27; Water Immersion, Nikon, Tokyo, Japan) at a scanning speed of one frame per second. An IR laser (Chameleon, Coherent, Palo Alto, CA, USA) was used to provide excitation at 820 nm. Fluorescence emission was collected on two detection channels: FF01-492/SP (400–492 nm) and FF03-525/50 (500–550 nm) (Semrock, New York, USA). The images provided in this article were obtained by merging these two detection channels.
Samples of the culture were collected and centrifuged at 8000× g for 5 min at 20 °C. The culture supernatant was removed, and the pellet was resuspended in PBS buffer (137 mM NaCl, 2.7 mM KCl, and 11.9 mM phosphate, pH 7.2) (ThermoFisher Scientific, Illkrich, France). Five microliters of cell suspension were placed between a glass slide and a coverslip for observation.
Samples of the culture were collected and centrifuged at 8000× g for 5 min at 20 °C. The culture supernatant was removed, and the pellet was resuspended in PBS buffer (137 mM NaCl, 2.7 mM KCl, and 11.9 mM phosphate, pH 7.2) (ThermoFisher Scientific, Illkrich, France). Five microliters of cell suspension were placed between a glass slide and a coverslip for observation.