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We demonstrate fluorescence imaging by two-photon excitation without scanningin biological specimens as previously described by Hwang and co-workers, butwith an increased field size and with framing rates of up to 100 Hz. Duringrecordings of synaptically-driven Ca$^{2+}$ events in primary rat neuronecultures loaded with the fluorescent Ca$^{2+}$ indicator Fluo-4 AM, we haveobserved greatly reduced photo-bleaching in comparison with single-photonexcitation. This method, which requires no costly additions to the microscope,promises to be useful for work where high time-resolution is required.

Widefield two-photon excitation without scanning: live cell microscopy with high time resolution and low photo-bleaching