MuVi SPIM multi-view light-sheet microscope

Bruker's Multi-View Selective-Plane Illumination Microscope (SPIM) has been developed based on years of experience and innovation of the Luxendo imaging system. MuVi SPIM has a modular design and is the fastest light-sheet system in the market. By using an acquisition unit exchange feature, the system allows for imaging of both live samples (LS) and cleared samples (CS).

MuVi SPIM enables imaging of cleared samples as well as high-speed imaging of dynamic processes in live samples and is compatible with any clearing technique.

• Best-in-class multiview imaging capabilities
  • Requires minimal striping artifacts to capture high-quality volumetric data and rapid biological processes.
• Multimodal modular concept
  • Enables cleared and live sample imaging with seamlessly interchangeable acquisition units (octagons).
• Advanced software and hardware integrations
  • Enables easy adjustment and precise configuration to meet the needs of current and future experiments.

Features

Best-in-class multiview imaging capabilities

Convenient multiview imaging

The system is available in various configurations and has a four-axis concept. This allows for

• High acquisition speed
• Long-term stability for imaging time series over days
• Simultaneous illumination and detection from two opposing directions without rotation
• Free rotation axis for optimal sample positioning and 360° view

Isotropic resolution with 360° illumination and detection

MuVi SPIM has a motorized sample holder that rotates samples through 360°. While in toto imaging does not require sample rotation, a single 90° rotation can improve image quality by providing isotropic resolution. The system is at least four times quicker than traditional SPIM.

Zebrafish expressing H2A::GFP acquired simultaneously from two opposing directions (left, right). Data fusion of the two views yields a high-quality in toto image of the sample (middle). Image Credit: Bruker Nano Surfaces and Metrology

Next-generation technology

The system has been developed based on Bruker’s years of experience and includes new features to reduce acquisition time, improve image quality, and enable artifact-free, high-resolution imaging of various samples.

New advanced features

Larger field of view (FOV) and superior axial resolution

Obstacles like pigments or cell nuclei can interfere with light during imaging, creating shadows and striping artifacts. The next-generation SPIM utilizes pivot-scan dual destriping to generate a uniform illumination profile with zero impact on acquisition speed.

Dual-destriping: Impressive elimination of striping artifacts

During the imaging process, any cell nuclei or pigments can interfere with light, leading to the generation of images with striping artifacts and shadows. The new SPIM enables dual destriping by using pivot scanning that allows for creation of a homogeneous illumination profile without changing the acquisition speed.

Left: Image collected without the destriping module showing striping artifacts. Center: Image collected with the destriping module showing successful removal of striping artifacts. Right: Merged image of both the left and right image. Image Credit: Bruker Nano Surfaces and Metrology

Improved signal-to-noise 

During imaging of opaque specimens or thicker samples, light scattering can blur the image and reduce contrast. MuVi SPIM has the following features to improve image contrast.

• Adjustable light-sheet thickness
• Line illumination for enhanced background suppression
• Scanned light sheet
• Robust aberration tolerance (even in complex samples)

TAG-lens: Light-sheet length control with fast axial scanning

Owing to advanced methods, ultra-fast varifocal lenses can be used to fine-tune the system’s optics. Luxendo SPIMs use tools driven by acoustic waves, called tunable acoustic gradient lenses (TAG lenses), for adaptive optics. These TAG lenses permit improved light-sheet management plus consistency while minimizing axial stretching.

Flexible live and cleared sample imaging options

Unique multimodal concept

MuVi SPIM has a multimodal and modular design, which allows researchers to select suitable capabilities and configuration depending on their experimental requirements. The system enables imaging of both cleared and live samples. This is possible by switching the optical core unit or the acquisition unit, the octagon, which holds the objective lenses. Besides the octagon, other modules, such as those for photomanipulation, can be exchanged or added easily. This flexibility offers many advantages in multi-user environments and in research.

MuVi SPIM LS - Designed for live, long-term imaging, with an optional photomanipulation module and complete environmental control. Image Credit: Bruker Nano Surfaces and Metrology

MuVi SPIM CS - Designed for cleared samples and supports any clearing solution, while ensuring optimal image quality. Image Credit: Bruker Nano Surfaces and Metrology

Transition between live sample and cleared sample system configurations in just minutes

• All the objective lenses and the sample chamber are placed on the octagon, the optical core unit.
• Different octagons can support different objective numerical apertures and magnifications and are compatible with objectives designed for either cleared or live sample imaging.
• The octagon can be easily replaced and removed while maintaining its optical alignment.

Applications

Pure live imaging

Native and natural environment

The system has a complete environmental control module to support imaging of different live specimens over hours and days without changing their biology. It allows for precise control of gas concentration (O₂, CO₂, N₂), temperature (20 to 37 °C), and humidity. The system thus finds application in

• Neurobiology and neurodevelopment
• Developmental biology
• Oncology
• Marine biology

Unparalleled imaging speed for in toto imaging

Using light-sheet technology, the system offers higher imaging speed than confocal microscopy. With low phototoxicity and high speed, it is possible to achieve in-depth yet gentle imaging of samples. This makes it applicable for fast biological dynamics such as blood flow or calcium imaging.

Advantageous photomanipulation

The capacity for spatiotemporal control, handling, and adjustment of biological functions remains central to uncovering the mechanisms involved. Adding one photo-manipulation (PM) component onto MuVi selective plane illumination microscopes (SPIM) enables trials including photoablation, cauterization, optogenetics, and fluorescence recovery after photobleaching (FRAP) using:

• Diffraction-limited illumination spot
• Continuous wave ultraviolet/visible (CW UV/VIS) to pulsed infrared (IR)
• Beam coupled into detection objective
• Flexible region-of-interest (ROI) generation (point, straight line, circle, freeform, and square)

Comprehensive cleared-sample imaging

Advanced cleared-sample applications

The system can be used alongside optical clearing techniques to enable non-destructive, detailed visualization of very large, in toto cleared specimens in minutes. This enables researchers to study

• Tumorigenesis
• 3D microstructure analysis of tissues
• Organ development
• Brain and central nervous system tissues

High flexibility of sample size and clearing method

MuVi SPIM can be used to image various cleared samples of different sizes, from 3D organoids to large tissue samples, up to the size of a cleared mouse brain. This is possible by:

• Different configurations of detection lenses to match the properties of the samples
• 15 × 15 × 20 mm scannable sample volume
• Novel optical design compatible with any kind of clearing solution

Innovative and easy sample handling

It is an innovative technique for mounting samples by reducing the user's exposure to clearing reagents. This is achieved by:

• Easy maintenance and cleaning of objectives and chamber
• Support of various mounting methods (e.g., hook, plate, pin, cuvette, etc.)
• Overhead 3D translation/rotation unit for sample mounting
• Fast objective exchange for easy transition between live-sample (LS) and cleared-sample (CS) imaging

Full environmental control

One of the key aspects of microscopy is the integrity and health of the samples. MuVi SPIM enables this by using an environmental control module to support imaging from hours to days. It has a large sample chamber for an immersion medium and has control over

• Gas concentration (CO₂, N₂, O₂,)
• Temperature 20–37 °C
• Humidity

Specifications

Configuration options

MuVi SPIM LS

Source: Bruker Nano Surfaces and Metrology

^* Calculation based on chip size.

MuVi SPIM CS

Source: Bruker Nano Surfaces and Metrology

^* Calculation based on chip size.

System comparison

Source: Bruker Nano Surfaces and Metrology

^* Compatible module

Software

All-in-one LuxBundle software

Intuitive design

Bruker's LuxBundle software reduces time while improving productivity by offering:

• Fully scriptable microscope control and post-processing through an open interface (e.g., Python or any other language), ready for custom "smart" microscopy
• Unified, user-friendly interface for acquisition, viewing, and post-processing
• File formats (.hdf5, .tiff, .ims) compatible with common image processing software: Imaris, BigDataViewer, Aivia, Fiji, Arivis, Matlab, Python, and Napari
• High reproducibility of experiments: All parameters are saved in the metadata and configurations can be saved for future experiments

Image Credit: Bruker Nano Surfaces and Metrology

Impressive image post processor

Bruker’s dual-view light-sheet microscopes can image a sample from different views/angles and generate images with multiple tiles. The LuxBundle software allows for 360° crisp images with improved quality to compensate for scattering and absorption. Features include:

• Tile stitching of hundreds of tiles for large samples
• Multi-color alignment
• Multi-view image fusion and deconvolution

Cleared mouse embryo labeled with methylene blue (cyan) and showing autofluorescence (magenta). Image composed of 12 tiles and 920 planes, all processed and stitched with LuxBundle. Image Credit: Montserrat Coll Lladó, European Molecular Biology Laboratory, Barcelona, Spain

3D data viewer

LuxBundle's integrated 3D data viewer enables researchers to analyze the complete dataset directly after acquisition. This gives users control over their data with key capabilities, including:

• Both raw and post-processed images
• The ability to turn tile stitching on or off
• Flexible options to draw and annotate regions and landmarks
• Fast viewing of multi-terabyte data sets