LysoSensor™ Green DND-189 - Special Packaging - FAQs

We recommend diluting the 1 mM stock solution to a working concentration of at least 1 µM using the growth medium or buffer of choice. To reduce potential artifacts from overloading, the concentration of dye should be kept as low as possible.

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Using a plate reader for measuring the pH of acidic organelles, such as lysosomes, with LysoSensor ratiometric probes, provides weak pH measurements, as discrete regions of interest cannot be selected for analysis. Therefore, there is no way of excluding extracellular fluorescence and little to no spatial or morphological information can be obtained. Instead, a readout of whole-cell fluorescence is provided, resulting in the average intraorganellar pH of all labeled organelles for the analyzed cell population. The is also the risk that a signal may be dominated by a few extremely bright, but essentially irrelevant cells, such as dead cells, or dye-labeled debris particles.

For more information about measuring luminal pH in organelles, please see the reference linked below:

Live-cell Microscopy and Fluorescence-based Measurement of Luminal pH in Intracellular Organelles

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For measuring the pH in acidic organelles, such as lysosomes, we recommend our LysoSensor ratio metric probes. Please see the links to the product page and descriptions below:

LysoSensor Yellow/Blue dextran, 10,000 MW, Anionic, Fixable (Cat. No. L22460):
LysoSensor Yellow/Blue dextran is a ratiometric probe that can be used to measure the pH of acidic organelles, such as lysosomes. This dextran conjugate is taken up by the cells through endocytosis. The LysoSensor dye produces blue fluorescence in neutral environments, but changes to yellow fluorescence in more acidic environments. The pKa of LysoSensor Yellow/Blue dextran is ~3.9.

LysoSensor Yellow/Blue DND-160 - Special Packaging (Cat. No. L7545):
LysoSensor Yellow/Blue DND-160 is a ratiometric probe that can be used to measure the pH of acidic organelles, such as lysosomes. The LysoSensor dye produces blue fluorescence in neutral environments, but changes to yellow fluorescence in more acidic environments. The pKa of LysoSensor Yellow/Blue DND-160 is ~4.2.

LysoSensor Blue DND-167 - Special Packaging (Cat. No. L7533):
LysoSensor Blue DND-167 can be used to measure the pH of acidic organelles, such as lysosomes. The LysoSensor dye becomes more fluorescent in acidic environments. The pKa of LysoSensor Blue DND-167 is ~5.1.

LysoSensor Green DND-189 - Special Packaging (Cat. No. L7535):
LysoSensor Green DND-189 can be used to measure the pH of acidic organelles, such as lysosomes. The LysoSensor dye becomes more fluorescent in acidic environments. The pKa of LysoSensor Green DND-189 is ~5.2.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.