UltraMicroscope II| Light sheet microscope | 3D Imaging

Benefits Features and Specifications Applications Support

Benefits

User friendly

An intuitive design, short training time for new users, and minimal staff support requirements make this light sheet microscope the perfect tool for imaging facilities.

Easy adaptation

Apply any clearing protocol and make use of the automated refractive index compensation to maintain high image quality.

High flexibility

Benefit from our objective lens series designed and optimized specifically for light sheet imaging. Our modular light sheet technology can be tailored to your needs.

From overview to cellular imaging

Explore cellular details in high resolution without losing sight of the big picture.

Features and Specifications

Zoom body configuration

Due to its easy handling, the Zoom body configuration is ideal for many applications. A large, easy-access sample chamber accommodates rodent organs and tumors, mouse embryos, insect larvae, or biopsies from large animals. An even illumination is achieved by six light sheets that illuminate the sample from both sides.

Super Plan configuration

The Super Plan Module is a motorized detection unit that can deliver an unprecented image quality and can be mounted directly to the focusing unit of existing UltraMicroscope II systems. Specially developed for light sheet microscopy, this setup maximizes imaging performance with the MI Plan objective series.

Technical features

Adjustable light sheets for best image quality

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The light sheet approximates a plane over a given horizontal range. This is where the light sheet is thinnest and where detection takes place. By setting the width, numerical aperture (NA), and Rayleigh length of illumination, users can optimize the shape of the light sheets for their specific sample size and imaging goals. A lower NA results in a broad field of view at the expense of a lower z-resolution and vice versa, with a full range of gradations in between.

Uniform axial resolution across large FOV

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Where high z-resolution and a large field of view (FOV) are essential, the UltraMicroscope II makes use of dynamic horizontal focus. A sequential series of high resolution images are taken across the desired FOV and automatically stitched together into a single high-quality image by our ImSpector image acquisition software.

Six light sheets for homogeneous illumination

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The 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.

Compatibility with all imaging solutions

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Most clearing reagents differ from protocol to protocol, and so does their optical properties. Light can travel differently in different imaging media, which can create distortions and thereby lower the image quality. To compensate for this, the UltraMicroscope II’s onboard software allows users to choose between various preloaded clearing and imaging solutions. This technology guarantees the perfect setting for every imaging media.

UltraMicroscope II specifications

Sheet optics
Illumination	Uni- and bidirectional
Number of light sheets	1–6
Thickness	4 µm–24 µm
Width	1 mm–20 mm
Numerical aperture	0.0135–0.135
Focus positioning	Dynamic
Refractive index matching	1.33–1.56
Detection optics	Zoom body configuration	Super Plan configuration
Objective lenses	2×	1.1×		4×	12×
Numerical aperture	0.5	0.1		0.35	0.53
FOV diagonal (5.5 Megapixel camera)	1.7–17.6 mm	≥33 mm		≥9.1 mm	≥3 mm
Zoom \| post-magnification	0.63×–6.3× (manual)	0.6×, 1×, 1.67×, and 2.5× (automated)
Total magnification	1.26×–12.6×	0.66×–2.75×		2.4×–10×	7.2×–30×
Working distance	5.6 mm (corrected), 10 mm	≤17 mm		≤16 mm	≤10.9 mm
Refractive index matching	1.33–1.56
Chromatic detection	Seven filters
Chromatic correction	Dynamic 400 nm–850 nm
Imaging chamber
Imaging solution	Aqueous buffers and organic solvents
Sample travel range (X, Y, Z)	1 cm, 1 cm, 1 cm
Sample size	µm range to cm range
Chamber size	72 mm × 74 mm × 35 mm
Light source
Laser module	Max. 5 laser lines (405, 488, 561, 639, 785 nm)*, 50 mW–100 mW per diode
Supercontinuum laser	Emission 460 nm–800 nm, 1 mW/nm–3 mW/nm
General information	Zoom body configuration		Super Plan configuration
Dimensions(W×H×D)	54 cm × 82 cm × 65 cm		54 cm × 73 cm × 65 cm
Weight	47 kg (w/o controller and laser)

^{*Other laser lines available upon request.}

Camera specifications

Detector
	5.5 Megapixel sCMOS camera			4.2 Megapixel sCMOS camera
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 size	6.5 µm × 6.5 µm			6.5 µm × 6.5 µm
Sensor size	16.6 mm × 14.0 mm 21.8 mm diagonal			13.3 mm × 13.3 mm 18.8 mm diagonal
Pixel well depth	30,000 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	200 MHz: 0.9 e- [1.2 rms]		560 MHz: 1.2 e- [1.6 rms]	216 MHz: 0.90 e- [1.1 rms]	540 MHz: 1.10 e- [1.3 rms]
Read noise [median] global shutter	200 MHz: 2.3 e- [2.5 rms]	560 MHz: 2.4 e- [2.6 rms]		-	-
Max readout rate	560 MHz			540 MHz
Max frame rates	Camera link: 100 fps USB 3.0: 53 fps			Camera link: 100 fps USB 3.0: 53 fps
Quantum efficiency	60%			82%

Upgrades - In vivo imaging module

View details

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 contro
Gas controller
Humidity module
Touch-screen operation board

Play video

Video

Zebrafish in vivo imaging

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

Resources

Brochures/posters

The UltraMicroscope II - Fast 3D imaging and analysis of entire biological systems

Scientific posters

In Vivo Labeling for Ex Vivo 3D Tracking of CAR T cells

References

Selected references

Culemann, S. et al. (2019) Locally renewing resident synovial macrophages provide a protective barrier for the joint. Nature
Esterházy, D. et al. (2019) Compartmentalized gut lymph node drainage dictates adaptive immune responses. Nature
Lowe, K. L. et al. (2015) Podoplanin and CLEC-2 drive cerebrovascular patterning and integrity during development. Blood 125(4): 3769-3777
Sjöstedt, E.. et al. (2020) An atlas of the protein-coding genes in the human, pig, and mouse brain. Science 367(6482)
Kimbrough, A. et al. (2020) Brain-wide functional architecture remodeling by alcohol dependence and abstinence. Proc. Natl. Acad. Sci. U.S.A.
Hägerling, R. et al. (2013) A novel multistep mechanism for initial lymphangiogenesis in mouse embryos based on ultramicroscopy. EMBO J. 32(5): 629-644
Weber, T. G. et al. (2014) Apoptosis Imaging for Monitoring DR5 Antibody Accumulation and Pharmacodynamics in Brain Tumors Noninvasively. Cancer Res. 74(7): 1913-1923
Belle, M. et al. (2017) Tridimensional Visualization and Analysis of Early Human Development. Cell 169(1): 161-173
Kirst, C. et al. (2020) Mapping the Fine-Scale Organization and Plasticity of the Brain Vasculature. Cell 180(4): 780-795
Renier, N. et al. (2014) iDISCO: A Simple, Rapid Method to Immunolabel Large Tissue Samples for Volume Imaging. Cell 159(4): 896-910
Renier, N. et al. (2016) Mapping of Brain Activity by Automated Volume Analysis of Immediate Early Genes. Cell 165(7): 1789-1802
Susaki, E. A. et al. (2014) Whole-brain imaging with single-cell resolution using chemical cocktails and computational analysis. Cell 157(3): 726-739
Tainaka, K. et al. (2014) Whole-body imaging with single-cell resolution by tissue decolorization. Cell 159(4): 911-924
Ye, L. et al. (2016) Wiring and Molecular Features of Prefrontal Ensembles Representing Distinct Experiences. Cell 165(7): 1776-1788
Pan, C. et al. (2016) Shrinkage-mediated imaging of entire organs and organisms using uDISCO. Nat. Methods 10: 859-867
Schmid, B. et al. (2019) 3Dscript: animating 3D/4D microscopy data using a natural-language-based syntax. Nat. Methods 16(4): 278-280
Todorov, M.I. et al. (2020) Machine learning analysis of whole mouse brain vasculature. Nat. Methods
Ertürk, A. et al. (2012) Three-dimensional imaging of solvent-cleared organs using 3DISCO. Nat. Protoc. 7(11): 1983-1995
Susaki, E. A. et al. (2015) Advanced CUBIC protocols for whole- brain and whole-body clearing and imaging. Nat. Protoc. 10(11): 1709-1727
Ueda, H.R. (2020) Tissue clearing and its applications in neuroscience. Nat. Rev. Neurosci. 21: 61-79
Kullmann, J. et al. (2020) Oxygen Tension and the VHL-Hif1α Pathway Determine Onset of Neuronal Polarization and Cerebellar Germinal Zone Exit. Neuron
Wan, Y. et al. (2019) Light-Sheet Microscopy and Its Potential for Understanding Developmental Processes Annu. Rev. Cell Dev. Biol. 35: 655-681
Liebmann, T. et al. (2016) Three-Dimensional Study of Alzheimer’s Disease Hallmarks Using the iDISCO Clearing Method Cell Rep 16(4): 1138-1152
Zundler, S. et al. (2017) Three-Dimensional Cross-Sectional Light-Sheet Microscopy Imaging of the Inflamed Mouse Gut. Gastroenterology 153(1): 898-900
Buglak, N. E. (2020) Light Sheet Fluorescence Microscopy as a New Method for Unbiased Three-Dimensional Analysis of Vascular Injury Cardiovasc. Res.
Nie, J. et al. (2019) Fast, 3D Isotropic Imaging of Whole Mouse Brain Using Multiangle‐Resolved Subvoxel SPIM Adv. Sci.
Claser, C. et al. (2019)
Lung endothelial cell antigen cross-presentation to CD8
⁺
T cells drives malaria-associated lung injury.
Nat Commun. 10(1): 5066
Levy, R.B et al. (2019) Circuit asymmetries underlie functional lateralization in the mouse auditory cortex. Nat Commun. 10(1): 2783
Lozovaya, N. et al. (2018) GABAergic inhibition in dual-transmission cholinergic and GABAergic striatal interneurons is abolished in Parkinson disease. Nat Commun. 9(1): 1422

Applications

Video

Immuno-oncology

Pancreatic carcinoma cell line with infiltrating CAR T cells

Reference list

Publications using UltraMicroscope light sheet imaging systems in cancer

Video

Neuroscience

Regeneration potential of neurons and axonal path finding

Reference list

Publications featuring UltraMicroscope light sheet imaging systems in neuroscience research

Support

Service where you need it

Fast and competent consultation by Miltenyi Biotec’s imaging specialists
Instrument training at MACS Academy Miltenyi Biotec facilities
Comprehensive service options provided by our globally distributed field service teams

UltraMicroscope II

Light sheet microscopy for 3D imaging and analysis of entire biological systems

Benefits

User friendly

Easy adaptation

High flexibility

From overview to cellular imaging

Features and Specifications

Zoom body configuration

Super Plan configuration

Technical features

Adjustable light sheets for best image quality

Uniform axial resolution across large FOV

Six light sheets for homogeneous illumination

Compatibility with all imaging solutions

UltraMicroscope II specifications

Upgrades - In vivo imaging module

Resources

Brochures/posters

Scientific posters

References

Selected references

Applications

Video

Immuno-oncology

Reference list

Publications using UltraMicroscope light sheet imaging systems in cancer

Video

Neuroscience

Reference list

Publications featuring UltraMicroscope light sheet imaging systems in neuroscience research

Support

Service where you need it