Standard Operating Procedure

 

CFSE Labeling

 

Hazards:

Caution - substance not yet fully tested. To our knowledge, the hazards of this material have not been thoroughly investigated. We recommend handling all chemicals with caution. (Taken from product MSDS)

 

Protection:

Wear gloves

 

Waste:

Drain

 

Spill Clean-Up:

 

Procedure:

• Product Name: 5-(and-6)-carboxyfluorescein diacetate, succinimidyl ester (5(6)-CFDA, SE; CFSE) *mixed isomers*
• Catalog Number: C1157
• Sold by Molecular Probes

 

1. Dilute 5mM CFSE 1:3 in HBSS (ex: 10ml CFSE into 20ml HBSS)
2. Resuspend cells to 10x10⁶ cells per ml (minimum of 1ml if fewer cells). Warm in H₂0 bath at 37°C for 10 min.
3. Add 1ml of diluted CFSE per 1ml of resuspended cells. Mix immediately by flicking or inversion.
4. Incubate in H₂0 bath at 37°C for 10 min.
5. Add ice cold RP10 to stop the reaction (min 10ml).
6. Wash with sterile PBS 1x.
7. Resuspend in sterile PBS inject.