Photometrics cool snap es ccd camera

Concentrated cells of WT and EYFP mutant strains from 2-day old cultures were deposited onto glass slides that were previously coated with 2% polyethyleneimine. Cells were imaged using a Nikon Eclipse 80i microscope equipped with a Photometrics Cool Snap ES CCD camera (Roper Scientific). Illumination was provided by a metal halide light source (X-Cite). Filter sets (Chroma) were as follows: YFP was detected using a 480/30 nm excitation filter, a 505 nm dichroic beam splitter, and a 535/40 nm emission filter. Chlorophyll fluorescence was detected using a 560/40 nm excitation filter, a 595 nm dichroic beam splitter, and a 630/60 nm emission filter. A 300 ms exposure time was used for imaging.

WT and eYFP containing mutant strains were concentrated 10-fold from mid log phase cultures. Samples were deposited onto glass slides that were coated with 2% polyethyleneimine. Cells were imaged using a Nikon Eclipse 80i microscope equipped with a Photometrics Cool Snap ES CCD camera (Roper Scientific). Filter sets (Chroma) were as follows: YFP was detected using a 480/30 nm excitation filter, a 505 nm dichroic beam splitter, and a 535/40 nm emission filter. Chlorophyll fluorescence was detected using a 560/40 nm excitation filter, a 595 nm dichroic beam splitter, and a 630/60 nm emission filter. A 100 ms exposure time was used for imaging chlorophyll fluorescence and a 1 s exposure time was used to image eYFP expression.