Fast 3D imaging of entire biological systems

UltraMicroscope II light sheet microscope

Features of the UltraMicroscope II

Adjustable light sheets

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The UltraMicroscope II has adjustable light sheets that allow the user to set width, numerical aperture (NA) and Rayleigh length. This unique feature provides highest flexibility. The user can choose settings that perfectly match their samples via the software.

Dynamic horizontal focus

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Custom-made optics form two triple light sheets that are focused into the sample. Z-resolution and contrast is best within the light sheet focus. To optimize this, the focus diameter and the focus length (Rayleigh length) can be adapted to the imaging conditions and will be optimized by software. Then, the UltraMicroscope II provides the dynamic horizontal focus that shifts the focus through the sample while imaging. In combination with advanced software algorithms, the UltraMicroscope II delivers a pin sharp image of the sample.

Six light sheets for homogeneous illumination

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LaVision BioTec’s UltraMicroscope II comes with 2×3  light sheets that excite the sample from both sides under slightly different angles. This means the fluorescence excitation is most homogeneous and artifacts like dark areas and stripes are minimized.

Multi-refractive index compensation

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Most clearing reagents differ from protocol to protocol and so does the refractive index. Running a system with different clearing or imaging solutions induces the necessity to correct for different refractive indices. The refractive index compensation of the UltraMicroscope II is utilized via the software interface. The user chooses between current clearing techniques such as CLARITY or Benzyl Ether. Water can also be selected for in vivo imaging. This technology guarantees the perfect setting for every imaging solution.

Applications

UltraMicroscope II specifications

Sheet optics 
IlluminationUni- & bidirectional
Number of light sheets3–6
Thickness4 µm–24 µm
Width1 mm–20 mm
Numerical aperture0.0135–0.135
Focus positioningDynamic
Refractive index matching1.33–1.56
Zoom microscope
 Zoom body setup Infinity corrected optics setup
Zoom Mono zoom -
Zoom ratio 0.63×–6.3× (1:10) -
Detection optics
 Zoom body setup Infinity corrected optics setup 
Objective lenses1.1×12×
Numerical aperture0.50.10.30.53
FOV diagonal [5.5 megapixel camera]1.7–17.6 mm19.7 mm5.4 mm1.8 mm
Total magnification
(objective lens + zoom ratio)
1.26×–12.6×(w/o zoom)(w/o zoom)(w/o zoom)
Working distance5.6 mm (corrected), 
10 mm
16 mm ,
17 mm
 
5.6 mm 8.5 mm, 
10 mm, 
10.9 mm
 
Refractive index matching1.33–1.56
Chromatic detectionSeven filters
Chromatic correctionDynamic 400 nm–850 nm
Imaging chamber 
Imaging solutionAqueous buffers and organic solvents
Sample travel range (X, Y, Z)1 cm, 1 cm, 1 cm
Sample sizeµm range to cm range
Light source 
Laser module              Max. 5 laser lines, 50 mW–100 mW per diode
Supercontinuum laserEmission 460 nm–800 nm, 1 mW/nm–3 mW/nm
General information 
 Zoom body setup Infinity corrected optics setup 
Dimensions 54 cm×82 cm×65 cm (W×H×D)54 cm×73 cm×65cm (W×H×D)
Weight 47 kg (w/o controller and laser)

Camera specifications

Detectors
 5.5 Megapixel sCMOS camera4.2 Megapixel sCMOScamera
Sensor type Front-illuminated scientific CMOS Front-illuminated scientific CMOS 
Active pixels (W×H) 2560×2160 (5.5 megapixel) 2048×2048 (4.2 megapixel) 
Pixel size6.5 µm2×6.5 µm26.5 µm2×6.5 µm2
Sensor size 16.6×14.0 mm
21.8 mm diagonal
 
13.3×13.3 mm 18.8 mm diagonal 
Pixel well depth30,000 e-30,000 e-
Pixel readout rate 200 MHz (100 MHz×2 sensor halves) 560 MHz (280 MHz×2 sensor halves) Slow Read 216 MHz (108 MHz×2 sensor halves) Fast Read 540 MHz (270 MHz×2 sensor halves) 
Readout modes Rolling shutter and true global shutter (snapshot) Rolling shutter and global clear  
Read noise [median]  Rolling shutter   Global shutter     Rolling shutter 
@ 200 MHz 0.9  e- [1.2 rms] 2.3 e- [2.5 rms] @ 216 MHz0.90 e- [1.1 rms]
@ 560 MHz 1.2 e- [1.6 rms] 2.4 e- [2.6 rms] @ 540 MHz1.10 e- [1.3 rms]
Max readout rate560 MHz 540 MHz 
Max frame ratesCamera Link: 100 fps
USB 3.0: 53 fps
Camera Link: 100 fps
USB 3.0: 53 fps
Quantum efficiency60% 82% 
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The UltraMicroscope II can be equipped with an incubation cuvette to ensure controlled environmental conditions. The temperature of the immersion medium as well as the atmosphere (CO₂/O₂) between medium and objective lens can be controlled using the okolab® environmental control system. All settings are controlled by a touch-screen. The heating element and the sample holder can be easily unmounted for autoclaving.

Content:

  • Incubation cuvette
  • Thermal controller with water bath
  • External temperature sensor installed directly at the sample for feedback temperature control
  • Gas controller
  • Humidity module
  • Touch-screen operation board
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Video

Zebrafish in vivo imaging

In vivo imaging of a zebrafish larvae with the UltraMicroscope II.