X cite fluorescent light source

For the quantification of nuclear enrichment of Hog1 terminal kinase in individual yeast cells as an indicator of pathway activation, we employed a strain in which a yellow-fluorescent protein (YFP) was integrated at the C-terminus of the endogenous Hog1 protein in Saccharomyces cerevisiae BY4741 yeast, using homologous DNA recombination.
Our imaging setup consisted of an inverted microscope, specifically the Nikon Eclipse Ti, which was equipped with several components, including perfect focus (Nikon), a 100× VC DIC lens (Nikon), a YFP fluorescent filter from Semrock, an X-cite fluorescent light source (Excelitas), and an Orca Flash 4v2 CMOS camera from Hamamatsu. The entire system was controlled via the Micro-Manager program.
To capture images, we conducted the following steps:
Utilizing the microscope’s xy-plane movement, we selected a field of view that exhibited an optimal density of yeast cells within a single z-plane. The z-focus was carefully adjusted to visualize the boundary of most cells, appearing as a white ring. Time-lapse images were recorded, encompassing both bright-field images, taken at intervals of every 10 seconds, and fluorescent channel images, acquired at intervals of every minute.