Figure 3
From: Efficient non-cytotoxic fluorescent staining of halophiles
Durability of MitoTracker staining during several growth and dilution cycles. (A) Schematic plot of OD600nm change during the experiment: 2-day exponential growth periods correspond to incubation time, while a 4-fold drop corresponds to the dilution. Particular time-points are labelled with lower-case Latin letters (a–j). (B) SNR decrease in the experiment. (C) (a) Microphotograph of cell culture at the day one; (d) two days after staining; (g) four days after staining and (j) six days after staining. Lower-case Latin letters in B and C correspond to the particular measurement time, shown in (A). Halobacteria were stained with MitoTracker Orange CMTMRos. The dye was washed out via centrifugation after staining. Cell cultures were incubated in a dye-free growth medium for 6 days with dilution every 2 days to maintain the active growth phase. The intensity of excitation laser was the same for all images. The detector gain was equal for images d,g,j, but it was reduced for image a to avoid saturation. The variation of the background in each image is due to different distances from the focal plane to the surface of the cover glass. Contrast stretching was applied to achieve the highest contrast of each image.
