Luxendo Light-Sheet Microscopes

TruLive3D Imager

The dual-sided illumination light-sheet microscope

TruLive3D Imager

The Bruker Luxendo TruLive3D Imager is optimized for fast 3D multi-sample volume imaging of delicate live specimens in their native 3D environment. Thus, it is particularly well-suited for time-lapse and high-throughput imaging of 3D spheroids, organoids, 3D cell cultures, and small embryos. Using an optical concept, with dual-sided illumination and single-lens detection from below, enables fast acquisition speed, high-resolution imaging, and minimal shadowing effects.

 

Latest technology advances allow for state-of-the-art uniform illumination and the optional addition of a photomanipulation module.

Imaging
Deep imaging and with high resolution
Inverted dual-sided illumination with our highest NA objective and with minimal shadowing effects.
Native and Natural
Environmental conditions
Environmental control for live-cell, long-term imaging of sensitive samples.
Multiplexing
Up to hundreds of samples
A sample holder for hundreds of samples and up to six different experimental conditions in one experiment.

Compact and Vibration-Free Design for Long-Term In Vivo Experiments

The TruLive3D Imager has a compact, robust and vibration-free design that provides maximal stability even during long-term and high-throughput experiments. Like all Bruker Luxendo's light-sheet solutions, the TruLive3D Imager is tailored to fit onto a lab bench, without the need for an air table. The proprietary piezo-crawler stages ensure longevity and precision for permanently accurate specimen positioning.

As the TruLive3D Imager is optimized for fast, long-term in vivo imaging, the system has advanced environmental control. This allows unparalleled imaging of experiments such as:

  • Spheroid and organoid cell-divisions, cell-interactions, and dynamics
  • 3D cell culture growth and drug responses
  • Small early embryo development, such as mouse, zebrafish, and drosophila
New Advanced Features    

Dual-Destriping: Impressive Elimination of Striping Artifacts

When imaging, obstacles like pigments or cell nuclei can interfere with light, leading to images with shadows and striping artifacts. The next-generation SPIM employs dual-destriping via pivot scanning, which generates a homogeneous illumination profile without compromising acquisition speed.

TAG-Lens: Light-Sheet Length Control with Fast Axial Scanning

Thanks to state-of-the-art technologies, ultra-fast varifocal lenses can be used to fine-tune the system's optics. Luxendo SPIMs use fluid-filled devices that are driven by an acoustic wave, called tunable acoustic gradient lenses (or TAG lenses), for adaptive optics. These TAG lenses allow better light-sheet control and uniformity while reducing axial stretching.

Next Generation Imaging

Deep and Crisp Imaging

Illumination and Detection

Using an inverted setup with dual-side illumination, single-lens detection, and dual-destriping, the TruLive3D Imager allows for minimal shadowing effects, uniform illumination, and deep sample penetration. In addition to advanced geometry, the TruLive3D Imager can achieve a high resolution.

While low phototoxicity is a key characteristic of light-sheet microscopy in general, the TruLive3D Imager takes sample-preserving imaging to the next step.

Native and Natural

An important goal in microscopy is sample health and integrity. To support this, the TruLive3D Imager is equipped with environmental control that supports long-term imaging (hours to days). In addition to a generous sample chamber for immersion medium, this includes control over:

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

Multiplexing

Large Sample Holder

To take organoid, spheroid, or 3D cell culture imaging to the next level, the TruLive3D Imager fits a large sample holder (length = 75 mm), which can accomodate hundreds of samples. Additionally, the new TruLive3D Dish series allows the user to separate the sample holder to student different experimental conditions at once. This setup makes the TruLive3D Imager ideal to study multiple samples over long periods of time and under different experimental conditions. An additional wide-field imaging option facilitates fast sample positioning.

Sample Embedding

Easy and intuitive sample embedding is key for enabling scientists to excel in their research. While the large sample holder (length = 75 mm) can accommodate hundreds of samples, the TruLive3D Dish series seamlessly integrates into the system and allows for testing of different conditions. Each TruLive3D Dish contains two sample spaces and three disposable dishes that extend the capacities of the system. Thus, a total of six experimental setups can be tested at once. For sample embedding, all holders use Fluorinated ethylene propylene (FEP) foil, which functions as a physical barrier and a "curved coverglass."

The FEP foil lining the holder serves dual functions as a “curved coverglass” and as a physical barrier between the immersion medium and sample medium. Samples can be raised directly on the foil or transferred, e.g., with a drop of matrigel. Importantly, only small volumes are needed for drug and compound testing, making the system highly effective. Using two LEDs and an overview camera allows easy sample localization and experiment setup through an overview camera looking from the side.

Inverted Setup for 3D Cell Culture Imaging        

Advanced Multiplex Acquisition

With the ability to study hundreds of samples at once and to perform long-term imaging for developmental and tissue studies, the TruLive 3D Imager provides a unique multiplex setup. To improve efficiency, we offer the option to partition the sample chamber, allowing researchers to study multiple drug treatments in parallel. This approach can save valuable time and resources, as time is often the scarcest commodity in research.

Innovative and Easy Sample Handling

The design of the TruLive 3D Imager allows for easy access from above and allows for swift immersion medium changes, which supports long-term imaging. Additionally, the system comes equipped with two LED illuminations which can be used for both sample positioning and experimental setup. These features can help streamline the imaging process and improve overall research productivity.

Long-Term Imaging

The TruLive 3D Imager is a state-of-the-art imaging system specifically designed for long-term imaging purposes. With its advanced features, the TruLive 3D Imager can acquire data over extended periods of hours and days all while ensuring the preservation of sample health and integrity. Its high quantum efficiencies of up to 82% provide excellent image quality, and its stage travel ranges of 66 x 3 x 3 mm allow for precise sample positioning. Additionally, this imager offers a wide range of final magnification options from 8x to 62.5x, making it a versatile tool for various research applications.

Imaging of Organoids and 3D Cell Cultures        

Organoids and Spheroids

Diabetes is a global health burden so understanding pancreas development and health is critical to learn more about the disease and treatment options. Using pancreatic spheres, one can study how cell fate choices and organ architecture play a role in pancreas health and disease.

 

Pancreatic Sphere

hESC-derived pancreatic spheres. Imaging on the TruLive3D Imager enables collecting information of several samples in one experiment. Visualization: Imaris (Bitplane)

Courtesy of:
Yung Hae Kim
Grapin-Botton Group, MPI-CBG
Dresden, Germany

 

 

Photomanipulation for Cell and Tissue Biology

Understanding how the immune system works and the various processes of immune defense is critical to understand health, disease, and vaccine efficacy. Often, immune system dynamics are impossible to measure with conventional approaches and need to be studied in vivo. This makes it a unique challenge for light-sheet microscopy.

Timelapse of cytokine dynamics in a tail wound assay. Pseudo-Brightfield and il1b:GFP (green) merged. Captured every 5 min for 12 hours.

Sample courtesy of: Dr. Elizabeth Jerison, Stanford University.

Tumorigenesis

Tumors are known to establish their own tissue ecosystem habitats. However, how tumors arise from a few malignant cells, how they interact with surrounding cells, and what this means for cancer progression is unknown. Having the ability to study tumorigenesis over time in a controlled system provides new insights into the decisions and processes of tumorigenesis.

Cell Biology in a Dish

The neurovascular unit is a critical component of the nervous system. However, the role of components such as glial cells is still not fully understood. Particularly brain glial cells, called astrocytes, play many different roles in neuron health and function. Thus so, understanding neuro-glial interactions is critical to understanding nervous system health. Reducing this system to "cell biology on a dish" adds control over the system, while enabling toxicological studies.

Specifications

Objective Tube Lens Magnification Pixel Size [nm] FOV [µm]
Nikon CFI APO LWD 25x 1.10 NA 100 12.5x 520 1065
  200 25x 260 532
  400 50x 130 266
Olympus XLUMPLFLN 16x 0.8 NA 100 8x 813 1664
  200 16x 406 832
  400 32x 203 416
  500 40x 163 333

Illumination Optics

  • Chromatic correction from 440 to 660 nm
  • Light-sheet generation by beam scanning
  • Flexible light-sheet thickness (2 µm to 6 µm)
  • Motorised ND filter set (90%, 10%, 1%) for high flexibility in using low laser power
  • Two Nikon CFI Plan Fluor 10x W 0.3 NA water immersion objective lens

Detection Optics

  • One Nikon CFI Apo 25x W 1.1 NA water immersion objective lens
  • Two spectral detection channels, each equipped with a fast filter wheel (10 positions and 50 ms switching time between adjacent positions) Filters adapted to the selected laser lines
  • Three motorised dichroic mirror positions for simultaneous dual channel acquisition
  • Motorised collar ring controlTwo 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

Accesories

TruLive3D Dishes

TrueLive3D Imager Sample Holder

Software

Luxendo's intuitive 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.

Electronics, Microscope Software and Computer

  • Embedded microscope software with an open communication interface: documented API, TCP/IP-based communication
  • Flexible GUI for interactive microscope control and experiment design
  • Computer with 256 GB RAM, Intel dual 8 core CPU
  • High-speed RAID controller for data streaming, 8×4 TB local storage in RAID 0
  • GeForce RTX 3070 graphics card
  • 10 Gbit/s on-board Ethernet port
  • 1 UHTV 43 inch display

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