MuVi SPIM

The multiview light-sheet microscope

MuVi SPIM LS
Key features
  • Unique 4-axis concept
  • 3D imaging of both large life specimens and cleared tissue
  • Compact, vibration-free and robust design
  • Resolution down to 280 nm in 3D
  • Browser-based user interface

Product overview

The MuVi SPIM is a horizontal setup designed to image large volumes very fast. The unique 4-axis concept with its two-sided illumination allows four combinations of illumination and detection.

This provides four orthogonal views of the specimen without the need for sample rotation, avoiding shadowing effects and facilitating long-term imaging. Combined with the sample mounting from below, this allows maximized acquisition speed and high precision of data fusion while preserving stability and integrity of the sample.

The MuVi SPIM capacities can be further extended to enable imaging of optically cleared samples and performing photomanipulation experiments (e.g. ablation, bleaching). For this purpose, the MuVi SPIM CS (Cleared Sample) and the MuVi SPIM PM (Photomanipulation) have been conceived.

Typical applications of the microscope include in toto imaging of animal models such as Zebrafish embryos, Drosophila larvae, marine biology samples, plant and root development, large cleared tissue samples. Adding the photomanipulation unit provides further flexibility, enabling optical interaction with the samples.

Device design

The MuVi SPIM’s compact, vibration-free and robust design provides maximal stability during your long-term experiments.

Tailored to fit your lab bench, this class 1 laser system does not require any air table or vibration compensation mechanism as all moving components are light-weighted and balanced with the motor technology. Maximal stability of focus and thermal conditions are also guaranteed. The proprietary piezo crawler stages ensure longevity and precision for a permanently accurate specimen positioning. Neither the images nor the natural growth behaviour of your sample are affected by the gentle image recording.

Illumination and Detection

The illumination optics comprise light-sheet generation by scanning a beam after passing a beam shaper unit for length and diameter adjustment.

The lenses project two aligned light-sheets from opposing directions on the sample. The expandable, alignment-free light source allows a range of up to six lasers (405, 445, 488, 515, 532, 561, 594, 642, and 685 nm with 40 mW each before fiber).

The detection optics include two high numerical aperture immersion objective lenses. The two spatial detection arms are equipped with 10 position high-speed filter wheels, and multiple spectral channels can be acquired sequentially. The robust sCMOS Orca Flash 4.0 V3 cameras from Hamamatsu are well-suited for experiments that require high detection efficiency, quantification and speed.

The sample chamber and all objective lenses are mounted on the octagon, the optical core unit that can be easily removed and replaced while maintaining its optical alignment. This makes the MuVi SPIM very flexible and user-friendly, a big advantage especially in multi-user environments.

Compared to other techniques, such as confocal laser scanning and spinning disc microscopy, more time points can be acquired for a deeper insight into dynamic biological processes and fast movement tracking, while reducing photodamaging effects, the major downside of the much higher illumination required in those techniques.

Electronics, microscope control and computer

LUXENDO's browser-based user interface offers a simple setup and execution of multidimensional experiments, while real-time control is handled by an embedded controller to ensure microsecond precision timing independent of the PC’s performance fluctuations.

Precise timing control of all connected devices is a prerequisite for reliable experimental outcomes. Full control of data streaming to storage as well as GPU-supported image processing further complements the overall performance.

Specifications

  • Electronics, microscope software & computer
    • Embedded microscope software with an open communication interface: documented API, TCP/IP-based communication
    • Browser-based GUI for interactive microscope control and experiment design
    • Computer with 128 GB RAM, Intel dual 8 core CPU
    • High-speed RAID controller for data streaming, 8 × 4 TB local storage in RAID O
    • GeForce GTX 1080 graphics card
    • 10 Gbit/s on-board Ethernet port
    • 1 UHTV 40 inch display

MuVi SPIM LS

(Live Sample)

The MuVi SPIM LS is the very first light-sheet microscopy system by LUXENDO.

Featuring a horizontal setup, it is designed to image large volumes of living objects very fast. The unique 4-axis concept with its two-sided illumination facilitates four orthogonal views of the sample without the need for rotation.

MuVi SPIM LS

Illumination and Detection

The MuVi SPIM can achieve a resolution down to 300 nm in 3D (at a wavelength of 500 nm), enabling live imaging, free of phototoxic effects.

Two Nikon CFI Plan Fluor 10x W 0.3 NA water immersion objective lenses project two aligned light-sheets from opposing directions on the sample. Detection includes two high numerical aperture Olympus 20x 1.0 NA or Nikon 16x 0.8 NA water immersion objective lenses. An additional magnification changer results in a total magnification that ranges from 12x to 33.3x.

Illumination Detection Effective Magnification Field of View Pixel size Optical Resolution
10x/0.3 NA Olympus 20x/1.0 NA 16.7x
22.2x
33.3x
800 µm
600 µm
400 µm
389 nm
293 nm
195 nm
300 nm
10x/0.3 NA Nikon 16x/0.8 NA 12.0x
16.0x
24.0x
1100 µm
830 µm
550 µm
542 nm
406 nm
271 nm
375 nm

Specifications

  • Illumination optics
    • Chromatic correction from 440 to 660 nm
    • Light-sheet generation by beam scanning
    • Flexible light-sheet thickness (2 µm to 8 µm)
    • 2 Nikon CFI Plan Fluor 10x W 0.3 NA water immersion objective lens
    • 2 identical illumination arms
  • Detection optics
    • Two Olympus 20x 1.0 NA or two Nikon 16x 0.8 NA water immersion objective lenses
    • 2 identical detection arms, each equipped with a fast filter wheel (10 positions and 50 ms switching time between adjacent positions)
    • Filters adapted to the selected laser lines
    • 2 high-speed sCMOS cameras Hamamatsu Orca Flash 4.0 V3
    • Maximum frame rate >80 fps at full frame (2048 × 2048 pixels of 6.5 µm × 6.5 µm size) and up to 500 fps at subframe cropping
    • Peak quantum efficiency (QE): 82% @ 560 nm

MuVi SPIM CS

(Cleared Sample)

The LUXENDO MuVi SPIM CS is an extension of the MuVi SPIM capabilities to enable 3D imaging of optically cleared samples.

The sample chamber, the illumination and the detection unit in the octagon are adapted to match the experimental needs in terms of sample size, refractive index, field-of-view and resolution, adding high flexibility to the system.

MuVi SPIM CS

Cleared Sample Module

The Cleared Sample Module is an add-on consisting of an exchangeable cleared sample octagon and an overhead sample positioning unit installed on the microscope’s top plate.

The octagon can be easily removed from the microscope for maintenance and cleaning of the sample chamber and the objective lenses that have been exposed to the mounting solution. The sample chamber is compatible with a broad range of solutions used in different clearing protocols. Available in two different sizes (max. volumes of 20 ml and 33 ml), the sample chamber can accommodate large samples such as mouse brains and other complete organs. Sample mounting from above enables easy sample access and optimized travel range suited for various mounting methods (e.g. hook, plate, pin or cuvette).

Flexible illumination and detection configurations leverage the MuVi SPIM CS to meet your application needs and allow imaging of a great variety of samples at high speed and high resolution.

Illumination and Detection

The MuVi SPIM CS (Cleared Sample) can achieve a resolution down to 300 nm in 3D (at a wavelength of 500 nm), enabling imaging of optically cleared samples.

Two Nikon 10x 0.3 NA air objective lenses project two aligned light-sheets from opposing directions on the sample. Detection includes high numerical aperture, multi-immersion objective lenses, with 20x and 10x magnification. An additional magnification changer results in a total magnification that ranges from 7.5x to 30x. You can choose the set of illumination and detection objective lenses according to the experimental needs.

Illumination Detection Effective Magnification Field of View Pixel size Optical Resolution
10x/0.3 NA Nikon 20x/1.0 NA 15.0x
20.0x
30.0x
890 µm
665 µm
445 µm
443 nm
325 nm
217 nm
300 nm
10x/0.3 NA Nikon 10x/0.5 NA 7.5x
10.0x
15.0x
1775 µm
1330 µm
890 µm
867 nm
650 nm
434 nm
600 nm

Specifications

  • Illumination optics
    • Chromatic correction from 440 to 660 nm
    • Light-sheet generation by beam scanning
    • Flexible light-sheet thickness (2 µm to 8 µm)
    • 2 Nikon 10x 0.3 NA air objective lenses
    • 2 identical illumination arms
  • Detection optics
    • Different combinations of lenses possible:

      • 1 Nikon 20x 1.0 NA 8 mm WD multi-immersion

      • 1 Nikon 10x 0.5 NA 5.5 mm WD

      • Combinations of 2 of them
    • 1 or 2 detection arms, each equipped with a fast filter wheel (10 positions and 50 ms switching time between adjacent positions)
    • Filters adapted to the selected laser lines
    • 2 high-speed sCMOS cameras Hamamatsu Orca Flash 4.0 V3
    • Maximum frame rate >80 fps at full frame (2048 × 2048 pixels of 6.5 µm × 6.5 µm size) and up to 500 fps at subframe cropping
    • Peak quantum efficiency (QE): 82% @ 560nm
  • Sample chamber
    • 1 sided 10x: 12 mm (lateral) × 19 mm (height) × 11 mm (depth)
    • 1 sided 20x: 12 mm (lateral) × 19 mm (height) × 16 mm (depth)
    • 2 sided 10x & 20x: 12 mm (lateral) × 19 mm (height) × 8 mm (depth)

MuVi SPIM PM

(Photomanipulation)

The LUXENDO MuVi SPIM PM is an extension of the MuVi SPIM capabilities to enable photomanipulation of your sample.

Whether you want to cut structures within cells and tissues (photoablation), photobleach fluorescence in selected regions of interest for FRAP or activate proteins for optogenetics, the MuVi SPIM PM module can be configured to meet your needs by choosing between three different types of lasers systems. It can be added to both MuVi SPIM LS and CS (photomanipulation only possible when LS octagon mounted).

MuVi SPIM PM

Illumination

The MuVi SPIM PM illumination beam is coupled into the right-side detection objective lens, creating a diffraction-limited illumination spot. Its size is determined by the properties of the detection objective lens.

A 2D scanner allows to freely position the illumination spot in 3D in the sample while it is being imaged – fully integrated in the acquisition software. Complex illumination regions (point, straight and freeform line, square) are either part of the experimental workflow or can be defined interactively on the fly while a 3D imaging experiment is carried out.

For different applications, the MuVi SPIM PM can be configured with different types of lasers, ranging from CW UV to pulsed IR lasers.

Detection objective Adressable FOV
Olympus 20x/1.0 NA 600 nm
Nikon 16x/0.8 NA 830 nm

Specifications

  • Illumination optics
    • Chromatic correction from 400 to 700 nm (UV/VIS) or from 780 to 1100 nm (IR)
    • Flexible ROI generation by beam scanning, available ROIs: point, straight and freeform line, square
    • Full software control
    • Diffraction-limited spot
    • Coupled into the right detection objective lens
    • Available lasers:
      • Pulsed IR laser 1030–1040 nm, 200 fs pulse length, 1.5 W, for photoablation
      • Up to two CW diode lasers, between 405 and 685 nm, up to 100 mW for photoactivation, photobleaching, optogenetics
      • CW or pulsed laser 780 nm, 100 fs pulse length, up to 500 mW, for photoactivation, optogenetics

Need help?

LUXENDO support

If you have any question regarding your microscopes functionality or you want to schedule a maintenance visit, then get in contact with LUXENDO’s support team. We will ensure to get the right person involved to answer your request.

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