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Two-photon glutamate uncaging

XT Xun Tu
RY Ryohei Yasuda
LC Lesley A. Colgan
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A second Ti-sapphire laser tuned at a wavelength of 720 nm was used to uncage 4-methoxy-7-nitroindolinyl-caged-l- glutamate (MNI-caged glutamate) in extracellular solution with a train of 4–8 ms, 2.8–3.0 mW pulses (30 times at 0.5 Hz) in a small region ~0.5 µm from the spine of interest as previously described14. Experiments were performed in Mg2+ fee artificial cerebral spinal fluid (ACSF; 127 mM NaCl, 2.5 mM KCl, 4 mM CaCl2, 25 mM NaHCO3, 1.25 mM NaH2PO4 and 25 mM glucose) containing 1 µM tetrodotoxin (TTX) and 4 mM MNI-caged L-glutamate aerated with 95% O2 and 5% CO2. Experiments were performed at room temperature.

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