5 ports Fluorescence Mini Cube - GCaMP and Optogenetic stimulation
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, the versions where LEDs are integrated as well reduce the number of optical connections and the system’s overall footprint.
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 - 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 57 mm x 63 mm x 26 mm Built-in Detectors Detector Sensitivity - 0.38 A/W @ 550 nm
- 0.60 A/W @ 960 nm (peak)
Amplification 3 levels : 1x, 10x, 100x Transimpedance gain - 2.0 E09 V / A (1x)
- 2.0 E10 V / A (10x)
- 2.0 E11 V / A (100x)
@ 550 nm
- 0.76 V / nW (1x)
- 7.60 V / nW (10x)
- 76.0 V / nW (100x)
@ 550 nm
- 7.2 nW (1x)
- 0.72 nW (10x)
- 72 pW (100x)
NEP < 12 fW/√Hz Bandwidth - DC : 0 - 1000 Hz
- AC : 30 - 1000 Hz
Electrical interface BNC Output Voltage 0.0 to 5.5 V Output impedance 50 Ω Sensor type Silicon photodiode Sensor Area 1 x 1 mm Input voltage 12 V DC (power supply included) Built-in LEDs Max Current 500 mA Emitter size 400 µm diameter - NA0.22 Max Output Power
(fibre 400 µm)
LED 400-410 nm
= 1.0 to 90 µW (adjustable)
LED 460-490 nm
= 1.5 to 120 µW (adjustable)
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