Actively helping customers, employees and the global community during the coronavirus SARS-CoV-2 outbreak.  Learn more >>

Is Calcein, AM toxic to cells?

Posted August 30, 2021


Answer

Calcein, AM is a cell-permeant dye principally used for determining cell viability. Once inside the cell, non-fluorescent Calcein, AM is hydrolyzed by non-specific intracellular esterases into a green fluorescent Calcein dye, which is negatively charged and well-retained within the cytoplasm. 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.

  • For fluorescence-based imaging we recommended the final concentration of Calcein, AM be 4 to 5 µM (with culture medium) or 2 µM (in HHBS buffer or buffer of your choice with culture medium removed).
  • For flow cytometry we recommend the final concentration of Calcein, AM be 500 nM to 1 µM.

The table below shows the range of available Calcein and CytoCalcein™ cell viability dyes, their excitation and emission maxima, and instrument specifications to assist you in selecting the best dye for determining cell viability.

Cat# Product Name Ex/Em (nm) Filter Set Channel Unit Size
22007 Calcein Blue, AM *CAS 168482-84-6* 351/439 DAPI Filter Set Pacific Blue Channel 1 mg
21908 Calcein UltraBlue™ AM 360/445 DAPI Filter Set Pacific Blue Channel 10 x 50 µg
22012 CytoCalcein™ Violet 450 *Excited at 405 nm* 405/444 DAPI Filter Set Pacific Blue Channel 1 mg
22013 CytoCalcein™ Violet 500 *Excited at 405 nm* 417/502 DAPI Filter Set
(Violet Filter Set recommended)
AmCyan Channel 1 mg
22002 Calcein, AM *CAS 148504-34-1* 500/520 FITC Filter Set FITC Channel 1 mg
22003 Calcein, AM *UltraPure grade* *CAS 148504-34-1* 500/520 FITC Filter Set FITC Channel 1 mg
22004 Calcein, AM *UltraPure grade* *CAS 148504-34-1* 500/520 FITC Filter Set FITC Channel 20 x 50 µg
21905 Calcein UltraGreen™ AM 496/524 FITC Filter Set FITC Channel 10 x 50 µg
22009 Calcein Orange™ diacetate 530/544 TRITC Filter Set PE Channel 1 mg
21900 Calcein Red™ AM 562/576 TRITC Filter Set PE Channel 1 mg
21902 Calcein Deep Red™ 642/663 Cy5 Filter Set APC Channel 1 mg
22011 Calcein Deep Red™ AM ester 642/663 Cy5 Filter Set APC Channel 1 mg
Additional resources

Calcein AM

Cell Viability Probes