Cell Explorer™ Fixable Live Cell Tracking Kit *Red Fluorescence*

Protocol summary

1. Prepare samples
2. Add 100 µL/well of 1X Track It™ Red working solution
3. Stain cells at 37°C for 15 minutes to 1 hour
4. Wash cells
5. Examine the specimen under microscope with Texas Red filter sets

Important notes
Thaw all the components to room temperature. Centrifuge component A briefly before opening.

1. Track It™ Red DMSO stock solution (500X):
Add 50 µL DMSO (Component C) into the vial of Component A. Note: The unused portion of the Track It™ Red stock solution should be aliquoted as a single used vials stored at -20 ^oC. Avoid light and repeated freeze/thaw cycles.

Dilute 500X Track It™ Red DMSO stock solution into Assay Buffer (Component B). For example, to get a 1X final concentration of Track It™ Red working solution for one 96-well microplate, dilute 20 µL of the Track It™ Red DMSO stock solution into 10 mL of Assay Buffer (Component B). Note: The final concentration of the Track It™ Red working solution should be empirically determined for different cell types and/or experimental conditions. It is recommended to test at the concentrations that are at least over a ten fold range. Note: The working solution should be prepared and used promptly. Keep from light.

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

1. Remove the cell medium, and add 100 µL/well (for 96 well plate) of 1X Track It™ Red working solution.
   
   
2. Incubate the cells in a 37°C, 5% CO₂ incubator for 15 minutes to 1 hour.
   
   
3. Wash cells with Hanks and 20 mM Hepes buffer (HHBS) or an appropriate buffer.
   
   
4. Fill the cell wells with growth medium or fix the cells (optional).
   
   
5. Analyze the cells using a fluorescence microscope with Texas Red filter sets or flow cytometer with 610/20 nm filter (PE-Texas Red channel).