HCS-FLIM

The goal for the end of the summer is to achieve a full working prototype system that will be capable of raster-scanning a fixed sample at a high speed and reconstruct a FLIM image. Improvements of the project would be:

  1. optical system: updated to include reference fiber and window galvo scanner
  2. reference fiber: display of reference spots on top of streak images, capable of real-time drift correction and time-axis calibration
  3. window galvo scanner: working in raster scanning mode, triggered by the main code
  4. main code: rebuilt to include communication with galvo drivers, new readout camera, real-time drift correction based on reference fiber

Beyond the summer, the project should shift to Spectral's facilities and Sunnybrook. At this point, the system would only require further characterization, extended programming, and a pilot study to validate the reconstructions. Aspects of the project that will be explored and further designed:

  1. pilot study: reconstruction of live cells expressing localized fluorescence and FRET standards to test validity
  2. optics improvements: explore changes to reference fiber, 2D-1D fiber array and streak camera input optics
  3. photon-counting efficiency: research and validate the photon requirements and optimal binning for lifetime estimation
  4. camera improvements: consideration of capabilities of EMCCD to replace the sCMOS readout

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