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Docket #: S13-139

Ultrafast Multifocal Multiphoton Microscope

Stanford researchers have developed an ultrafast multi-foci two-photon microscope system that aims at 1 kHz full frame rate with 500x500 ?m2 field of view (FOV). It utilizes a 2D foci-array pattern and 1D scanning mechanism to achieve full FOV excitation coverage. A customized high-speed intensified CMOS camera is used for parallel detection of fluorescence signals from all laser foci. A custom image processing algorithm is then performed to reconstruct a complete image frame from its multiple sub-frames while correcting for optical cross talk between nearby foci. This invention advances state-of-the-art in vivo microscopy to match the recent progress of fluorescent voltage indicators with higher imaging speeds and large FOV.

Related Microscopy Technologies from the Schnitzer Lab
Stanford docket S13-155: "Robotic system for simultaneous multiple-area optical microscopy of a three-dimensional sample"
Stanford docket S13-154: "Very large curved field of view two-photon multi-foci raster scanning-microscope"

Figure


Overview of the main frame of the optomechanical design of the system

Applications

  • General two-photon / Confocal imaging
  • Electrophysiology imaging
  • In-vitro / In-vivo imaging

Advantages

  • Ultrafast imaging speed – from 10-20 Hz to 1kHz full frame
  • Large FOV
  • 2D tilted foci array pattern and 1D scanning mechanism
  • Customized intensified camera system as the photon collection device for two-photon imaging
  • Customized deconvolution algorithm to recover image quality
  • Conventional two-photon / one-photon imaging capability included
  • Advances state-of-the-art in vivo microscopy to match the recent progress of fluorescent voltage indicators with higher imaging speeds and large FOV

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