Fig. 5. LysoSensor Green DND-189 is a vital dye that fluoresces in the acidic
compartments of the yeast cell. (A) Comparison of LysoSensor Green DND-189 and
Quinacrine staining. Wild-type cells were stained separately with LysoSensor
Green DND-189 and Quinacrine as described in Materials and methods. (B)
Wild-type cells were incubated at 0 °C for 30 min with 30 µmol
l-1 FM 4-64 in YPD medium buffered with 100 mmol l-1
Hepes, pH 7.6. The cells were harvested by centrifugation, washed once with
the same buffer, and resuspended in 0.35 ml of fresh medium containing 4
µmol l-1 LysoSensor Green DND-189. Cells were then incubated for
5 min, harvested by centrifugation, washed and resuspended in YPD medium.
Portions were removed for microscopy and photographed at 25 min (top) and 45
min (bottom). Top left of each panel, FM 4-64 fluorescence; top right, the
same cells in visible light; bottom left, LysoSensor Green DND-189
fluorescence; bottom right, the double-labeled image. The images were
recorded, merged and processed using the Zeiss LSM Image Browser.
