Cell Explorer™ Live Cell Labeling Kit *Green Fluorescence*

Protocol summary

1. Prepare cells in growth medium
2. Remove the medium
3. Add Calcein Green™ working solution (100 µL/well for 96-well plates or 25 µL/well for 384-well plates)
4. Incubate the cells at 37 ^oC for 30 - 60 minutes
5. Wash the cells
6. Examine the specimen under under fluorescence microscope with FITC filter (Ex/Em = 490/525 nm)

Important notes
Thaw all the components at room temperature before starting the experiment.

1. Calcein Green™ stock solution:
Add 20 µL of DMSO into the vial of Calcein Green™ (Component A) and mix well to make Calcein Green™ stock solution. Note: 20 µL of Calcein Green™ stock solution is enough for 1 plate. For storage, seal tubes tightly. Note: Unused Calcein Green™ stock solution can be aliquoted and stored at < -20 ^oC for more than one month if the tubes are sealed tightly. Avoid repeated freeze-thaw cycles and protect from light.

Add 20 µL of Calcein Green™ stock solution into 10 mL of HHBS (Component B) and mix well to make Calcein Green™ working solution. Note: Protect from light.  

For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html

1. Remove the growth medium from the cell plates.
   
   
2. Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of Calcein Green™ working solution into the cell plate.
   
   
3. Incubate the cells in a 37°C, 5% CO₂ incubator for 30 to 60 minutes. 
   
   
4. Wash the cells with HHBS (Component B), and add growth medium or HHBS back to the cells.
   
   
5. Image the cells using a fluorescence microscope with FITC filter (Ex/Em = 490/525 nm).