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Bundle Fluorescence with targeted optogenetics

Bundle Fluorescence with targeted optogenetics
Bundle Fluorescence with targeted optogenetics


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  • The Bundle-Imaging Fiber Photometry system with targeted optogenetics (BFTO) enables multi-site measurements through High-density Fiber-optic Patch Cords (HDP). These patch cords are available in either square or hexagonal patterns of 7, 9 or 19 fibers connected to the sample port, allowing combined optogenetics and photometry recording .


    Build-in LEDs combine Isosbestic & Functional GCaMP excitations with Red Fluorophore excitation. Isosbestic and functional fluorescent signals are recorded with the same image sensor by interleaving the excitations. Both the LED driver and the CMOS detector are integrated within the system. The stimulation pattern can vary in type, number and emitted wavelengths. For this purpose, a second similar high-density fiber optic patch cord is connected to the optogenetic port and fans out into separate connectors. Then, up to 19 independent optogenetic stimulation inputs can be connected depending on the needs of the experiment (one input per fiber connected to the sample port).
    The BFTO system with targeted optogenetics supports the following fluorophore and opsin combinations:

     

    Fluorophores

    Green Fluorophore (GCAMP, dLight)

    Exc 460-490nm / Em 500-540nm

    Red Fluorophore (RCAMP, Jrgeco1A, dTomato)

    Exc 555-570nm / Em 580-600nm

    Opsins

    ChR2 (450 nm)

    N/A1

    Sequential2

    NpHR/Chrimson (590 nm)

    Simultaneous or Sequential

    N/A1

    Jaws (632 nm)

    Simultaneous or Sequential

    Simultaneous or Sequential

    1Opsin stimulation is within the excitation or emission range of the fluorophore.
    2ChR2 stimulation at 450nm generates optical fiber autofluorescence signal
    3ArchT stimulation at 560 nm may cause crosstalk in the 500-540 detection band.


    • Wavelength range 350 to 1100 nm
      Field of view Ø3 mm
      Objective NA 0.40
      Maximum number of sites*
      		 - 7x core 400 µm NA0.37
      - 9x core 400 µm NA0.37
      - 19x core 200 µm NA0.37
      LEDs Excitation Uniformity 90% over FOV
      Optogenetics Excitation Uniformity 65% over FOV
      Optogenetics Crosstalk between fibers >41 dB
      Optical fiber compatibility - Core diameter 200, or 400 µm
      - NA 0.37
      Spectral bandwidth - Excitation #1 : 405-410 or 410-415 nm
      - Excitation #2 : 460-490 nm
      - Excitation #3 : 555-570 nm
      - Emission #1 : 500-540 nm
      - Emission #2 : 580-600 nm
      Optical filter attenuation > OD 5 outside band
      Optical fiber connector HD7, HD9 or HD19
      Dimensions 220mm x 220mm x 59mm
      SENSOR
      Type CMOS image sensor
      Resolution 1024x1024 px
      Pixel size

      5.86 µm x 5.86 µm
      Frame rate up to 60 Hz
      Computer interface USB 3.0
      * could be limited by patchcord manufacturing
    • Connectorized LEDs
        Drawing
      Connectorized LEDs & LED Drivers
        User Manual