title: Multifocal multiphoton microscopy (MMM) at a frame rate beyond 600 Hz.
abstract: We introduce a multiphoton microscope for high-speed three-dimensional (3D) fluorescence imaging. The system combines parallel illumination by a multifocal multiphoton microscope (MMM) with parallel detection via a segmented high-sensitivity charge-couple device (CCD) camera. The instrument consists of a Ti-sapphire laser illuminating a microlens array that projects 36 foci onto the focal plane. The foci are scanned using a resonance scanner and imaged with a custom-made CCD camera. The MMM increases the imaging speed by parallelizing the illumination; the CCD camera can operate at a frame rate of 1428 Hz while maintaining a low read noise of 11 electrons per pixel by dividing its chip into 16 independent segments for parallelized readout. We image fluorescent specimens at a frame rate of 640 Hz. The calcium wave of fluo3 labeled cardiac myocytes is measured by imaging the spontaneous contraction of the cells in a 0.625 second sequence movie, consisting of 400 single images.
authors: Karsten
authors: K
authors: Bahlmann
authors: Peter T
authors: PT
authors: So
authors: Michael
authors: M
authors: Kirber
authors: Robert
authors: R
authors: Reich
authors: Bernard
authors: B
authors: Kosicki
authors: William
authors: W
authors: McGonagle
authors: Karl
authors: K
authors: Bellve