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Two-photon excitation of channelrhodopsin-2 at saturation.

ABSTRACT: We demonstrate that channelrhodopsin-2 (CR), a light-gated ion channel that is conventionally activated by using visible-light excitation, can also be activated by using IR two-photon excitation (TPE). An empirical estimate of CR's two-photon absorption cross-section at lambda = 920 nm is presented, with a value (260 +/- 20 GM) indicating that TPE stimulation of CR photocurrents is not typically limited by intrinsic molecular excitability [1 GM = 10(-50)(cm4 s)/photon]. By using direct physiological measurements of CR photocurrents and a model of ground-state depletion, we evaluate how saturation of CR's current-conducting state influences the spatial resolution of focused TPE photostimulation, and how photocurrents stimulated by using low-power scanning TPE temporally summate. We show that TPE, like visible-light excitation, can be used to stimulate action potentials in cultured CR-expressing neurons.

SUBMITTER: Rickgauer JP

PROVIDER: S-EPMC2736443 | biostudies-literature | 2009 Sep

REPOSITORIES: biostudies-literature

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Two-photon excitation of channelrhodopsin-2 at saturation.

Rickgauer John Peter JP Tank David W DW

Proceedings of the National Academy of Sciences of the United States of America 20090814 35

We demonstrate that channelrhodopsin-2 (CR), a light-gated ion channel that is conventionally activated by using visible-light excitation, can also be activated by using IR two-photon excitation (TPE). An empirical estimate of CR's two-photon absorption cross-section at lambda = 920 nm is presented, with a value (260 +/- 20 GM) indicating that TPE stimulation of CR photocurrents is not typically limited by intrinsic molecular excitability [1 GM = 10(-50)(cm4 s)/photon]. By using direct physiolog ...[more]

PMID: 19706471

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