Is calcein AM toxic to cells?
Is calcein AM 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 does calcein work?
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 dissolve calcein am?
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.
What is the function of the AM 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 long does calcein AM last in cells?
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.
How long does calcein AM last?
We use Calcein AM to allow us to take images of neuronal processes for analysis of their length and branching. 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.
Does calcein AM stain dead cells?
Dead cells either do not stain with calcein due to lack of esterase activity, or fail to retain calcein in the cytoplasm due to compromised plasma membrane integrity.
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.
Does calcein AM method assay the total cellular labile iron pool or only fraction of it?
In the present study we show that the calcein-AM method does not capture lysosomal low-mass iron and, therefore, that the method seriously underestimates total cellular labile iron.
Can calcein AM Be Fixed?
Can I fix the cells after labeling and retain the staining? This is not recommended. Neither Calcein nor ethidium homodimer-1 bind to any cellular components upon fixation. There is no guarantee that the dyes will be retained upon fixation or any subsequent wash steps.
What is the purpose of using calcein AM?
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.
What is the function of calcein in iron chelation?
Calcein, a fluorescent iron chelator, has been widely used to monitor the labile iron pool. When the non-fluorescent acetoxymethyl ester moiety (calcein-AM), enters cells, it is immediately cleaved by cytosolic esterases to its cell-impermeable, fluorescent calcein form.
Can calcein measure labile iron?
High levels of labile iron have been shown to promote oxidative stress. As this pool has such profound effects upon cellular iron homeostasis, there has been great interest in the development of methods to measure labile iron. Calcein, a fluorescent iron chelator, has been widely used to monitor the labile iron pool.
Does calcein perturb cellular iron homeostasis?
Furthermore, we have found that the size of the labile iron pool as measured by calcein was proportional to the amount of calcein loaded in HeLa and K562 cell lines. These findings suggest that calcein may be able to perturb cellular iron homeostasis, and may not accurately reflect the size of the labile iron pool.
