Dye Solution

Recipe

MATERIALS

HEPES

KCl

NaCl

Oregon Green 488 BAPTA-1 (OGB-1) acetoxymethyl (AM)

Pluronic F-127, 20% solution in DMSO (dimethyl sulfoxide) (Molecular Probes/Invitrogen)

Filter, 0.2-µm pore (e.g., Semadeni, Ostermundigen, Switzerland)

Sonicator

Ultrasonic bath (e.g., Elma Transsonic T310)

1. Prepare HEPES-buffered solution (125 mm NaCl, 2.5 mm KCl, 10 mm HEPES).
2. Prepare 50 µg OGB-1 AM in 5 µL of Pluronic solution.
3. Sonicate the OGB-1 AM solution for 15 min.
4. Dilute the OGB-1 AM solution in 28 µL of HEPES-buffered solution.
5. Filter through a 0.2-µm filter.
6. Mix for another 15 min with an ultrasonic bath.
- This solution can be stored at −20°C. We recommend using within 2 mo.