C9100 23b emccd camera

Live-cell imaging was accomplished using a PerkinElmer Ultraview spinning disk microscope equipped with a Nikon Apochromat TIRF 100X 1.49 numerical aperture objective and a Hamamatsu C9100-23B EMCCD camera, and 405, 488, and 561 nm lasers were used in the experiments (www.perkinelmer.com). Images were acquired by Volocity (www.perkinelmer.com). To observe yeast cells, stack images containing 11 planes at 0.5 μm spacing were acquired. Yeast cells were observed on slides coated with EMM agarose (3%) pads.
Microscopic data were analyzed with Fiji ImageJ 1.52 (www.imagej.nih.gov) and MetaMorph 7.7 (www.moleculardevices.com). Graphs were created by KaleidaGraph 4.5 (www.synergy.com). Normality of data was determined by OriginPro (version 2021b) (www.originlab.com), and statistical analysis was performed by KaleidaGraph 4.5. The illustration graph in Figure 6 was created using graphs generated with BioRender.com.

All imaging experiments were carried out with a PerkinElmer Ultraview Spinning Disk confocal microscope equipped with a Nikon Apochromat TIRF 100X 1.49NA objective and a Hamamatasu C9100-23B EMCCD camera (PerkinElmer). Images were acquired at room temperature with log-phase cells sandwiched between an EMM5S agarose pad and a coverslip. For high-temporal resolution imaging, stack images containing three planes with 0.5 μm spacing were taken every 5 sec by Volocity (PerkinElmer) unless otherwise specified; for single time-point imaging, stack images containing 11 planes with 0.5 μm spacing were acquired.

All imaging data were acquired at room temperature by Volocity on a PerkinElmer ultraview spinning-disk confocal microscope equipped with a Nikon Apochromat TIRF 100X 1.49NA objective and a Hamamatasu C9100–23B EMCCD camera (www.perkinelmer.com). For maximum projection images, stack images containing 11 planes at 0.5 μm spacing were taken. Data were analyzed with MetaMorph 7.7 (www.moleculardevices.com) and ImageJ 1.52 (NIH, Bethesda, MD, USA). Graphs were generated and statistical analysis was performed with KaleidaGraph 4.5 (www.synergy.com). Diagram illustrations were created with BioRender.com.