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| Multiphoton laser unit |
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In this setup the IR laser is coupled to the second scanner (for stimulation) - to be used for pinpoint stimulation during confocal imaging. It still can be used for MPE imaging with NDD detection. |
| Multiphoton near-infrared pulsed laser |
• Mode-locked Ti:sapphire laser (femtosecond) with integrated "negative chirp" pulse pre-compensation system
• Possible lasers
Spectra-Physics:
MaiTai BB DeepSee (710nm - 990nm)
MaiTai HP DeepSee (690nm - 1040nm)
Coherent:
Chameleon Vision I (690nm - 1040nm)
Chameleon Vision II (680nm - 1080nm)
• Includes laser power unit and water-cooled circulating chiller for Ti:sapphire laser |
| Shutter/attenuator |
AOM |
| Integration optics |
• For stable and highly efficient integration of IR laser into the scan unit
• Motorized keplerian beam expander
• Laser safety measures implemented via protective covers |
| Single photon laser unit |
Visible light laser |
• Diode lasers: 405nm (50mW), 440nm (25mW), 473nm (15mW), 635mW (20mW)
• DPSS laser: 559nm (15mW)
• Multi Ar laser: 457nm, 488nm, 515nm (total power 30mW)
• HeNe laser: 543nm (1mW) |
| AOTF laser combiner |
• Visible light laser platform with implemented AOTF system, ultra-fast intensity control for every individual laser line
• Laser intensity modulation: AOTF (0.1 - 100% in 0.1% increments)
• Laser turn-off during the retrace period
• Connected to scanner via single-mode fibre
• Equipped with feedback mechanism to ensure constant laser intensity |
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| Scanning unit for imaging |
Laser ports |
• First port: visible-light laser introduction by fibre
• Second port: free, but prepared for introduction of multiphoton near-infrared pulsed laser via direct coupling by air
• Third port: free |
| Optical components |
• Dichromatic mirrors for fluorescence excitation (both single- and multiphoton excitation)
• High efficiency coating of lenses for visible and IR light
• High-performance filter for fluorescence emission and as IR-cut filters |
| Scanning method |
• 2 galvanometer scanning mirrors
• Pixel size: 64 x 64 - 4096 x 4096 pixels |
| Scanning Speed |
• Standard mode: time per pixel variable ~ 2 µs - 200µs
• Standard mode: 1 frame/sec (512x512) and lower
• High-speed scanning mode: 16 frames/sec (256 x 256) |
| Scanning modes |
• Dimensions: Time, Z, (wavelength) and any combination thereof
• Line scan: straight line (includes rotation), free line, point XY scan |
| Field Number |
18 |
| Zoom size |
1 - 50x (adjustable in 0.1x increments) |
| Scanning unit for stimulation |
Laser ports |
Multiphoton near-infrared pulsed laser port via direct coupling by air |
| Optical components |
Exchangeable dichromatic mirrors for combination of laser light via imaging scanner with laser light via laser scanner for stimulation |
| Scanning method |
• 2 galvanometer scanning mirrors
• Pixel size: 64 x 64 - 4096 x 4096 pixels |
| Scanning Speed |
Standard mode: time per pixel variable ~ 2 µs - 200µs |
| Scanning modes |
• Full frame, regions of interest
• Line scan: straight line (includes rotation), free line
• Point scan: single or multiple points
• Tornado scan (for higher stimulation efficiency) |
| Field Number |
18 |
| Zoom size |
1 - 50x (adjustable in 0.1x increments) |
| Non-descanned detection (NDD) unit |
Detection modules |
• In the reflective pathway with two or four channels
• In the transmitted pathway (e.g. for SHG) with two channels (only with upright microscope) |
| Detectors |
PMT detectors with analog integration and hybrid phtoton counting modes |
| Separation of fluorescence |
• All detectors in equal distance to objective
• 32mm diameter filters and dichroic mirrors |
| Confocal detection unit |
Pinhole |
Single motorised pinhole, adjustable in 0.5 µm increments |
| Configuration |
• Fluorescence detection: three or four channels
• Transmitted light detection: one channel
• Additional port for exiting fluorescence (e.g. fibre coupled) |
| Detectors |
PMT detectors with analog integration and hybrid photon counting modes |
| Fluorescence separation with filters |
Dichromatic mirrors and highly efficient filters used on all channels |
| Spectral fluorescence separation |
• Channels 1 and 2 provided with independent grating and slit
• Selectable wavelength range: 400 - 800 nm
• Wavelength resolution: 2 nm
• Wavelength selection bandwidth: 1nm - 100nm
• Wavelength switching speed: 100 nm/ms |
| Transmitted light unit |
Coupling to microscope |
• External detector unit integrating transmitted laser light detector and transmitted light source
• Motorised switching |
| Contrast methods |
Standard transmitted light, DIC (not possible in combination with IR laser), phase contrast (not recommended with laser scanning) |
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| Z-drive |
Drive |
Motorised focus module inside microscope frame |
| Minimum step size |
0.01 µm |
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| Microscopes |
Motorised microscopes |
• Upright microscope BX61WI (with moving objective nosepiece)
• Inverted microscope IX81 |
| System control |
Operating system |
• Windows XP Professional (English version)
• Windows Vista |
| PC specifications |
Current PC specifications on request |
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| Monitors |
• Standard: 30''
• Optional two 21'' |
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| Software |
FV10-ASW 2.0 (or later) |
| Vibration isolation table |
Vibration control |
Pressured air (compressor, bottle or house supply) |
| Table size |
1.5m x 1.5m (W x D) or larger |
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| Table top |
Whole pattern with 25mm and M6 thread |
| Required installation environment |
Room temperature |
20°C — 25°C ± 1°C |
| Humidity |
60% or less |
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| Dust level |
Class 1000 |
| Power source |
AC100-120/220-240V 60VA, continuous (24-hour) power supply required for laser |
Optimal for in vivo observation
