Cell Explorer™ Live Cell Labeling Kit *Blue Fluorescence*

Protocol summary

1. Prepare cells in growth medium
2. Remove the medium
3. Add Calcein UltraBlue™ 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 minutes to 2 hours
5. Wash the cells
6. Examine the specimen under under fluorescence microscope with DAPI filter (Ex/Em = 360/445 nm)

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

1. Calcein UltraBlue™ stock solution:
Add 20 µL of DMSO into the vial of Calcein UltraBlue™ (Component A) and mix well to make Calcein UltraBlue™ stock solution. Protect from light. Note: 10 µL of Calcein UltraBlue™ stock solution is enough for 1 plate. For storage, seal tubes tightly.

Add 10 µL of Calcein UltraBlue™ stock solution into 10 mL of HHBS (Component B) and mix well to make Calcein UltraBlue™ 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 UltraBlue™ working solution into the cell plate.
   
   
3. Incubate the cells in a 37°C, 5% CO₂ incubator for 30 minutes to 2 hours.
   
   
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 DAPI filter (Ex/Em = 360/445 nm).