Oregon Green BAPTA-1 AM Dye Mixture
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1. Take an appropriate amount of Oregon Green BAPTA-1 AM (OGB-1). Be careful to keep it dry.
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2. Raise the temperature of the OGB-1 by holding the vial in your gloved hand.
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3. Prepare a solution of dimethylsulfoxide (DMSO) containing 20% (w/v) Pluronic and keep it at 60°C for 30 min. Dissolve the OGB-1 in 4 µL of DMSO/20% Pluronic. The OGB-1 solution should look slightly yellowish.
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4. Vortex the mixture for 1 min.
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6. Add 1 µL of 2 mm Alexa 594 or 1 µL of 2 mm sulforhodamine 101.
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7. Vortex the mixture for 1 min.
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8. Seal the top of the tube with Parafilm.
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9. Sonicate the mixture for 5 min.
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10. Filter the dye mixture through a 0.45-μm mesh using a centrifuge (e.g., Millipore Ultrafree-MC HV Centrifugal filter (UFC30HV00).
- © 2014 Cold Spring Harbor Laboratory Press
