5 ports Fluorescence Mini Cube - GCaMP and Optogenetic stimulation

5 ports Fluorescence Mini Cube - GCaMP and Optogenetic stimulation
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  • Dual excitation band fiber photometry combined with optogenetics use a Fluorescence Mini Cube with 5 ports: one for 460-490 nm GCaMP excitation, one for 405 or 415 nm isosbestic excitation, one for 500-540 nm window of isosbestic and functional fluorescence detection, one for 580-650 nm opsin silencing light, and one for the sample under the test. The cube has dichroic mirrors to combine isosbestic and fluorescence excitations, isolate the fluorescence emission, and let opsin stimulation light through. At the same time, narrow bandpass filters limit the excitation fluorescence spectrum.

    E stands for fluorescence excitation, IE for isosbestic excitation, F for fluorescence, O for opsin stimulation light, and S for the sample port on the cube's schematic.

    The separation of functional and isosbestic fluorescence signals of GCaMP is possible with modulated light sources. The Intensity Adjustment Ring on the 460-490 nm LED light source reduces its power below the LED driver's range if needed for low-power fiber photometry experiments.

    Compared to Fluorescence Mini Cubes with only FC connectors on all ports, the ones with integrated photodetectors provide a 100% better signal-to-noise ratio (30% signal increase and a 40% noise reduction). Moreover, in the latest version of Mini Cubes, LEDs and LED drivers are integrated as well to reduce the number of connections and the system’s overall footprint. In his version, it is also possible to replace the excitation filter (to adjust the isosbestic excitation from 405 nm to 410 nm for instance) and decrease the excitation power by adding attenuating filters (for low power photometry).

  • Mini Cube
    Wavelength range 350 to 1100 nm
    Optical fiber compatibility - Core diameter 200 or 400 µm
    - NA 0.37 to 0.57
    Spectral bandwidth

    GCaMP :
    - Isosbestic Excitation 400-410 nm
    - Excitation 460-490 nm
    - Emission 500-540 nm
    Optogenetics : 580-680 nm

    Optical filter attenuation > OD 5 outside band
    Dimensions - FMC: 82 x 59 x 26 mm
    - iFMC: 90 x 84 x 50 mm
    - ilFMC: 107 x 109 x 52 mm
    Built-in Detectors iFMC ilFMC
    Detector Sensitivity - 0.38 A/W @ 550 nm
    - 0.60 A/W @ 960 nm (peak)
    - 0.38 A/W @ 550 nm
    - 0.60 A/W @ 960 nm (peak)
    Amplification 3 levels : 1x, 10x, 100x 2 levels : 1x, 10x
    Transimpedance gain - 2.0 E09 V / A (1x)
    - 2.0 E10 V / A (10x)
    - 2.0 E11 V / A (100x)
    - 2.0 E09 V / A (1x)
    - 2.0 E10 V / A (10x)
    Conversion gain
    @ 550 nm
    - 0.76 V / nW (1x)
    - 7.60 V / nW (10x)
    - 76.0 V / nW (100x)
    - 0.76 V / nW (1x)
    - 7.60 V / nW (10x)
    Saturation level
    @ 550 nm
    - 7.2 nW (1x)
    - 0.72 nW (10x)
    - 72 pW (100x)
    - 7.2 nW (1x)
    - 0.72 nW (10x)
    NEP < 12 fW/√Hz < 12 fW/√Hz
    Bandwidth - DC : 0 - 1000 Hz
    - AC : 30 - 1000 Hz
    - DC : 0 - 1000 Hz
    - AC : 30 - 1000 Hz
    Electrical interface BNC BNC
    Output Voltage 0.0 to 5.5 V 0.0 to 5.5 V
    Output impedance 50 Ω 50 Ω
    Sensor type Silicon photodiode Silicon photodiode
    Sensor Area 1 x 1 mm 1 x 1 mm
    Input voltage 12 V DC (power supply included) 12 V DC (power supply included)
    Built-in LEDs (ilFMC)
    Max Current 500 mA
    Emitter size 400  µm diameter - NA0.22
    Max Output Power
    (fibre 400 µm)
    LED 400-410 nm
    = 60 µW (at 200 mA, CW)*
    LED 460-490 nm
    = 130 µW (at 200 mA, CW)*
    * with the ND filter T=10% included.
  • Drawing

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