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What is LysoSensor?

What is LysoSensor?

Description. 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.

Is LysoTracker deep red fixable?

LysoTracker dye is a cell permeable dye which stains acidic organelles in live cells. Generally, it can’t be used in already fixed tissues as fixation with alcohol or aldehydes changes pH gradient required to stain acidic compartments.

What is a LysoTracker?

LysoTracker® Deep Red is a deep red-fluorescent dye for labeling and tracking acidic organelles in live cells. Features of all LysoTracker® probes include: • High selectivity for acidic organelles. • Effective labeling of live cells at nanomolar concentrations.

How does LysoTracker red work?

Due to its hydrophobic nature (Log P = 2.1 5), LysoTracker Red enters cells via simple diffusion. Remarkably, owing to the acidic pH of the lysosome, upon entry into the lysosome, LysoTracker Red becomes protonated and hence is markedly sequestrated in lysosomes.

How does LysoSensor green work?

LysoSensor Green is a weak base that accumulates in acidic organelles in response to both pH gradients and absolute organellar pH, and whose fluorescence is increased upon protonation.

Can LysoTracker be fixed?

LysoTracker-stained cells can also be fixed in paraformaldehyde followed by mounting in Vectashield mounting medium without significant loss of fluorescent signal (Cao et al., 2006).

What does MitoTracker red stain?

Description. MitoTracker Red CMXRos is a red-fluorescent dye that stains mitochondria in live cells and its accumulation is dependent upon membrane potential. The dye is well-retained after aldehyde fixation.

How does LysoSensor work?

The LysoSensor™ family of fluorescent dyes (ThermoFisher Scientific) provides a means for fluorescence-based measurement of lysosomal pH. In living cells, the fluorescent dye produces yellow fluorescence in acidic environments, such as lysosomes, whereas it produces blue fluorescence in neutral environments.

Is DAPI a Counterstain?

DAPI is a popular nuclear counterstain for use in multicolor fluorescent techniques. Its blue fluorescence stands out in vivid contrast to green, yellow, or red fluorescent probes of other structures. When used according to our protocols, DAPI stains nuclei specifically, with little or no cytoplasmic labeling.

What is MitoTimer?

MitoTimer is a novel fluorescent reporter protein that can reveal new insights into mitochondrial dynamics within cells. Coupled with organelle flow cytometry, it offers new opportunities to investigate mitochondrial subpopulations by biochemical or proteomic methods.

Which is better, acridine orange or lysotracker red?

Acridine orange makes it possible to distinguish acidic vesicles with different internal pH but is fairly phototoxic and can cause spectacular bursts of the dye-loaded vesicles. LysoTracker Red is less phototoxic but its rapid photobleaching limits the range of useful applications considerably.

What should the dye concentration be for the lysotracker?

For the LysoTracker® probes, we recommend working concentrations of 50–75 nM and for the LysoSensor™ probes at least 1 µM. To reduce potenti al artifacts from overloading, the concentration of dye should be kept as low as possible.

How are lysosensor probes used in fluorescent staining?

Choose a LysoTracker or LysoSensor probe for fluorescent staining of lysosomes in cells. Summary of our LysoTracker and LysoSensor probes—Table 12.3 | Thermo Fisher Scientific – US

Is the base of lysotracker red partially protonated?

A molecule of LysoTracker Red consists of a fluorophore linked to a weak base that is only partially protonated at neutral pH. Similarly to AO, it is likely that the molecule is protonated on entry into an acidic organelle and the protonated form is far less likely to cross the membrane again.