Basic Calcein AM Cell Viability Kit

This kit allows researchers to easily and simultaneously differentiate between live and dead cells within a single sample. Samples can be analyzed using a flow cytometer, fluorescence plate reader, or fluorescent microscope.

Figure 1. Live cells exhibiting green fluorescence (right of histogram) are easily distinguished from dead cells (unstained, left of histogram). Jurkat cells were either mock-treated (A) or killed by exposure to 90% ethanol (B). Cells were stained with Calcein AM, then analyzed in a flow cytometer with a FL-1 99% attenuation filter. Most (93.0%) live cells stained positive with Calcein AM (A, right), while 0.0% of the dead cells stained positive (B, right). Data courtesy of Dr. K. Strandberg.
Figure 2. Mock-treated Jurkat suspension cells were stained with Calcein AM to detect live cells. The majority of the cells imaged were considered to be live and healthy. Panel A reveals green fluorescence-stained live cells. Panel B shows a corresponding differential interference contrast (DIC) image, which reveals cell morphology. Microscope images were obtained using a Nikon Eclipse 90i microscope with a Hamamatsu Flash 4.0 camera. Data courtesy of Dr. Kristi Strandberg (ICT 226:95).
Jurkat suspension cells were mock-treated (black) or killed by exposure to 90% ethanol (red). A mix of the live and dead cells is shown in gray. Cells were stained with 0, 1, and 10 µM Calcein AM to detect live cells and analyzed using a fluorescence plate reader. The fluorescence signal in live cells increased with increasing concentration of Calcein AM. Dead cells had little or no fluorescence. Data courtesy of Dr. Strandberg (ICT 227:1-2).
Figure 1. Live cells exhibiting green fluorescence (right of histogram) are easily distinguished from dead cells (unstained, left of histogram). Jurkat cells were either mock-treated (A) or killed by exposure to 90% ethanol (B). Cells were stained with Calcein AM, then analyzed in a flow cytometer with a FL-1 99% attenuation filter. Most (93.0%) live cells stained positive with Calcein AM (A, right), while 0.0% of the dead cells stained positive (B, right). Data courtesy of Dr. K. Strandberg.
Figure 1. Live cells exhibiting green fluorescence (right of histogram) are easily distinguished from dead cells (unstained, left of histogram). Jurkat cells were either mock-treated (A) or killed by exposure to 90% ethanol (B). Cells were stained with Calcein AM, then analyzed in a flow cytometer with a FL-1 99% attenuation filter. Most (93.0%) live cells stained positive with Calcein AM (A, right), while 0.0% of the dead cells stained positive (B, right). Data courtesy of Dr. K. Strandberg.
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Product Manual
Safety Data Sheet

SKU: 9153

Size: 25 - 250 Tests
Price:
$242.00
Ships: 1-2 business days
  1. Prepare samples and controls.
  2. Dilute 10X Cellular Assay Buffer 1:10 with diH2O.
  3. Reconstitute Calcein AM with 50 µL DMSO to prepare 2 mM stock solution.
  4. Dilute 2 mM Calcein AM stock solution 1:5 by adding 200 µL diH2O or PBS, forming a 400 µM stock solution.
  5. Stain with Calcein AM at a concentration between 1-10 µM. The ideal staining concentration can vary based on cell line, application, etc., and should be determined by the end user. The recommended sample size is 0.4 mL.
  6. Add Calcein AM to each sample and mix gently. For example, to stain a 0.4 mL sized sample with 10 µM Calcein AM, add 10 µL of the 400 µM stock solution.
  7. Incubate approximately 1 hour.
  8. Analzye with a flow cytometer, fluorescence microscope, or fluorescence plate reader. Calcein AM excites at 494 nm and emits at 520 nm.
  • Calcein AM Reagent, 1 vial, #6696
  • Hoechst 33342, 1 vial, #639
  • 10X Cellular Assay Buffer, 60 mL, 1 bottle, #6695
  • Kit Manual

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