What is the function of the acetoxymethyl ester group on calcein?

The non-fluorescent acetomethoxy derivate of calcein (calcein AM, AM = acetoxymethyl) is used in biology as it can be transported through the cellular membrane into live cells, which makes it useful for testing of cell viability and for short-term labeling of cells.

How do you dissolve calcein?

Calcein AM, in contrast, is susceptible to hydrolysis. Therefore, it should be dissolved in organic solvents like DMSO with a concentration of approximately 10 mg/mL.

How long does calcein AM stain last?

Over the short term (12-24 hours) Calcein AM does not seem to kill neurons. Some minimal neuronal death is seen by 48 hrs of culture with Calcein AM and this is increased by 72 hrs culture.

Is calcein AM fixable?

Calcein, AM diffuses into cells, the ‘AM’ moiety is cleaved by cellular esterases, and then the dye molecules are observed in the cytoplasm without binding to anything. This gives a ‘whole cell’ stain. It also means that the dyes are not crosslinked with aldehyde-based fixation and thus will be lost upon fixation.

What is calcein used for?

Calcein AM is a cell-permeant dye that can be used to determine cell viability in most eukaryotic cells. In live cells the nonfluorescent calcein AM is converted to a green-fluorescent calcein after acetoxymethyl ester hydrolysis by intracellular esterases.

How do you make a calcein indicator?

Dissolve 0.2 g of bis-N,N-di(carhoxymethyl) amino methyl- fiuorescein (G. Frederick Smith Chem- ical Company, No. 222) in 25 ml of 0.1 N sodium hydroxide and dilute to 100 ml with water.

How do you dilute calcein?

dilute the 2 mM Calcein AM stock solution 1:5 by adding 200 µL PBS. This creates a 400 µM Calcein AM solution. If staining at a final concentration of 10 µM, then this solution is ready to use.

Is calcein am light sensitive?

Calcein AM Dye is supplied at ambient temperature. Upon arrival store at -20°C. The product when stored as directed is stable for 1 year. light as Calcien AM dye is light sensitive.

Is calcein toxic to cells?

Calcein, AM has been used for studies of enzymatic activity, cell membrane integrity, and long-term cell tracking due to its low cellular toxicity. It is important to note that for certain cell lines and experimental conditions, relatively high concentrations of Calcein, AM can be toxic to cells.

How do you take calcein?

Add 100 µl of working stock solution of Calcein AM dye and incubate the cells for 30 minutes or 1hr in incubator (5%CO2, 37°C). NOTE: For most cell types, 30 minutes incubation is adequate.