Confotec CARS is a multifunctional multichannel 3D scanning laser microscope-spectrometer, combining in one system:
- CARS scanning microscope
- Raman/luminescent scanning confocal microscope
- conventional scanning confocal laser microscope
Notable for its high resolution and high-speed scanning, the system allows of performing real time noninvasive analysis of biological samples (cells and components of living cells) with high spatial resolution.
Multifunctional - Confotec CARS combines:
Multi-channel - five channels for simultaneous high-speed measurements:
Advantages of CARS method
Features High spatial resolution:
Wide spectral range:
High spectral resolution:
Simultaneous / multifunctional analysis:
Three types of CARS measurements:
Five independent speed channels for simultaneous detection up to four 2D and 3D images:
Three scanning modes:
Highly-precise calibration over the wavelengths: better than ±0,002nm due to an embedded calibration lamp as a source of reference lines for automated operative calibration of the monochromator-spectrograph. |
Polysterene balls of various diameter (F-CARS, 3045 cm-1) |
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3D CARS image of liquid crystal 8CB structure on resonant frequency 2236 cm-1 |
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Multimodal CARS/TPEF image of growing cancer cells HeLa: DNA/RNA/Proteins/Lipids colored |
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Selective imaging of MIA-PaCa pancreatic human cancer cell for resonance 2845 cm -1 C-H bound (C-H aliphatic stretch). Selective lipids visualization. |
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3D image of a cell |
Common specifications
Spatial XYZ resolution of CARS signals (Objective lens: 60х, NA=1.2, watter immersion): |
< 0.7 μm |
Spatial resolution of Raman signals (excitation wavelength 633 nm; objective lens: 60х, NA=1.2, watter immersion): |
XY: < 300 nm Z: < 700 nm |
Spectral detectable range: |
CARS signals: 985 – 5000 cm-1 |
Spectral resolution: | CARS signals: 7-8 cm-1 Raman signals: 0.25 cm-1(grating 75 l/mm Echelle); 0.6 cm-1 (grating 1800 g/mm) |
Scanning range (fast scanning mode, 60x lens) | XY: 225 х 225 μm Z: 80 μm |
Control and automation: | Fully motorized |
Optical microscope
Type: | inverted |
Model: | Nikon Ti-U |
Stage: | motorized |
- travel range: | 114 х 75 mm |
- accuracy (for 1 mm traveling): | 0.06 μm |
- XY repeatability: | ± 1 μm |
- minimal step: | 0.02 μm |
Micro- objective lens: | 60 х NA-1.2 water immersion 20 x NA-0.45 |
Z-scanner: | |
- type: | piezo scanner |
- lens movement range: | 80 μm |
- minimal step: | 50 nm |
- Reapetability: | < 6 nm |
Illuminator for reflection mode: | Halogen lamp 100 W |
Illuminator for transmission mode: | LED |
Laser input port: | motorized triple turret |
High resolution digital video camera: | |
- type: | digital color CCD camera |
- sensor: | 1/2", 2048 x 1536 pixeles |
- ADC: | 10 bit, 12 frames per min |
CARS excitation laser system
Single module optical parametric laser system consisting of: | |
Picosecond solit state Nd:YVO4 laser | |
Output wavelength: | 1064 nm |
Pulse duration: | 6.5 ps |
Output power: | 6 W |
Pulse repetition rate: | 85 MHz |
Mode composition: | TEM00 |
Beam quality (М2): | < 1.5 |
Beam diameter: | 2 mm |
Tunable SOPO (Synchronously pumped Optical Parametrical Oscillator) |
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Wavelength tuning range: | 690 - 990 nm |
Output power: | > 400 mW (at 800 nm) |
Pulse duration: | 5 - 6 ps |
Pulse repetition rate: | 85 MHz |
Polarization: | horizontal |
Mode composition: | TEM00 |
Beam quality (М2): | < 1.2 |
Beam diameter: | 2 mm |
Built-in delay tuning optical line |
Laser for Raman spectroscopy
Laser type: | cw HeNe |
Wavelength: | 632.8 nm |
Average output power: | > 10 mW |
Polarization: | linear |
Beam diameter: | 0.65 mm |
Beam divergence: | < 1.24 mrad |
Laser beam convergence and appointment unit
Laser shutter: | 3 pieces (Nd:YVO4 laser, SOPO and HeNe laser) |
Polarizer: | 2 pieces (Nd:YVO4 laser and SOPO) |
Wavelength plate λ/2 | 2 pieces (Nd:YVO4 laser and HeNe laser) |
Variable telescope: | 3 pices (Nd:YVO4 laser, SOPO and HeNe laser) |
Main optical unit
Optimized optics for spectral range: | 400 - 1100 nm |
Polarization analyzer: | Glan-Teylor prism |
Laser beam attenuator: | veriable neutral density filter 0 - 3D |
Pinhole lens positioner: | three-axis (X, Y, Z) |
CARS, Raman, fluorescence filters positioner: | five- position |
Posotioner of dihroic mirrors: | six- position |
Imaging monochromator/spectrograph MS5004i
Focal length: | 520 mm |
F number (input): | 9.8 |
Horizontal magnification: | 1.0 |
Vertical magnification: | 1.0 |
Vertical spatial resolution: | < 20 μm |
Flat field size: | 28 х 5 mm |
Stray light ( 20 nm from 633 nm laser line) |
1 х 10-5 |
Difraction gratings unit: | 4-position motorized grating turret |
Spectral resolution (wavelength 633 nm, CCD pixel size: 12x12 μm) |
0.01 nm (Echelle grating 75 l/mm) |
Entrance spectral slit | motorized confocal pinhole, continuously adjustable from 0 to 1.5 mm |
Output spectral slit: | motorized, width from 0 to 2 mm (continuously adjustable) |
Ports: | 1 input, 2 output |
Detector (Raman, E-CARS, fast measurements) | Hamamatsu Photosensor module H7844 (with TE cooling) |
Monochromator for fluorescence measurements (integrated with MS5004i)
Focal length: | 100 mm |
Input spectral slit: | motorized confocal pinhole |
Output spectral slit: | fixed, width 4.2 mm |
Difraction grating: | 600 l/mm |
Spectral range: | 400 - 920 nm |
Linear reciprocal dispersion: | 13 nm/mm |
Detector | Hamamatsu Photosensor module H7844 (with TE cooling) |
Spectral CCD camera (Raman, E-CARS)
Type: | digital CCD camera |
Photosensor: | back-thinned CCD sensor 2048 х 122 pixels |
Pixel size: | 12 х 12 μm |
Photosensitive area: | 24.576 х 1.464 mm (length х heigh) |
Spectral sensitivity range: | from 200 to 1100 nm |
Cooling with temperature stabilization: | two-stage Peltier (TE) cooling down to – 45 °С |
ADC digitization: | 16 bit |
Sensitivity: | 1 photon for 1 ADC count (at 650 nm) |
Dinamic range: | at least 10 000 |
Fast scanning unit (X, Y)
Scanners: | Galvano-mirror scanners (X, Y) |
Scanning model: | raster high speed and start-stop mode |
Positioning accuracy: | < 30 μm |
Scanning area: | 225 μm х 225 μm (objective lens 60Х) |
Full frame scanning speed: | 4 s/frame: 1000 х 1000 points |
Laser confocal microscope unit (Reflected)
Prepinhole lens positioner: | three -axis (X, Y, Z) |
Confocal pinhole: | motorized confocal pinhole, continuously adjustable from 0 to 1.5 mm |
Detector: | Hamamatsu Photosensor module H6780-01 |
F-CARS and transmitted signal registration system
Polarization alanyzer: | Glan-Teylor prism |
F-CARS filters positioner: | 4-position, motorized |
F-CARS signal registration detector: | Hamamatsu Photosensor module H7844 (with TE cooling) |
Transmitted signal registration detector: | Hamamatsu Photosensor module H6780-01 |
Registration and processing control unit
Number of registration channels: | 5 |
Number of simultaneously registered channels: | up to 5 |
PC connection interface: | Ethernet 100 Base-T (TCP/IP protocol) |
Optical table
All components of the system based on the optical plate with the pneumatic vibrated insulating legs.