How do you store rhodamine?

After first opening of the bottle, the contents can be used up to the stated expiry date when stored at +5 °C to +30 °C. The bottles must be kept tightly closed at all times.

Can a MitoTracker be fixed?

For MitoTracker® Red CMXRos, we have found that fixing with 3.7% formaldehyde in complete growth medium at 37°C for 15 minutes works well for endothelial cells.

How long does MitoTracker last?

approximately 48 hours
I want to know about the stability of the mitotracker dyes. For how long is the fluorescent intensity maintained? Is it possible to add the dye, then perform the experiment and then visualize the cells. The experiment will last approximately 48 hours.

How toxic is rhodamine?

For Rhodamine B, the effective and lethal concentration (EC50 and LC50) –causing 50% toxicity were in the range of 14–24 mg/L. For Rhodamine WT, no statistically significant effects were observed (p<0.05) at the tsted concentrations (up to 91, 100 and 200 mg/L for algae, crustaceans and fish embryos, respectively).

What is mitochondrial depolarisation?

mitochondrial depolarization The process in which the potential difference across the mitochondrial membrane is reduced from its steady state level.

What does MitoTracker red bind to?

However, MitoTracker is chemically reactive, linking to thiol groups in the mitochondria. The dye becomes permanently bound to the mitochondria, and thus remains after the cell dies or is fixed. In addition, it can be used in experiments in which multiple labeling diminishes mitochondrial function.

Why is MitoTracker added to live cells?

The mitochondria can be labelled with mitotracker because this reagent present electronic mitochondrial potential affinity. If you fix the cells before use mitotracker will not work because you don’t have electronic mitochondrial potential. If you have already the fixed cells, you need use antibody.

Why is rhodamine 123 used in this assay?

The potentiometric fluorescent dye rhodamine 123 (Molecular Probes, Eugene, OR) is used in this assay to provide a relative assessment of electrochemical potential across the mitochondrial membrane.

How is mitochondrial loading with rhodamine 123 performed?

Mitochondrial loading with rhodamine 123 is performed by washing cultures in situ once with PBS, and then incubating the cultures with 5 μ g/ml rhodamine 123 in serum-free medium (prepared from 2 mg/ml stock in ethanol). Incubation time is based on kinetic analysis of rhodamine 123 loading, which is typically 30 minutes in astroglia.

Why do we use fluorescein and rhodamine 123 as anionic substrates?

Fluorescein and rhodamine 123 are used as anionic and cationic substrates because they are cheap, safe to handle, and sensitively determined by fluorescence spectrometry. The process requires energy in the form of ATP, which can serve as another measurement endpoint (i.e., released phosphates from the hydrolysis of ATP).

What is the pathophysiology of DHR 123 oxidation?

DHR 123 enters the cell and gets oxidized by ROS to rhodamine 123 (RH 123) and accumulates in the mitochondria. The ability of RH 123 to compartmentalize in the mitochondria on oxidation is due to the formation of the cationic rhodamine 123 iminium cation (in resonance with xanthylium salt), which exhibits green fluorescence (Fig. 4.12 ).