InVi SPIM

The light-sheet microscope optimized for long-term 3D fluorescence imaging

InVi SPIM
Key features
  • Close-to-natural environmental conditions for sensitive samples
  • Compact, vibration-free and robust design
  • Resolution down to 255 nm in xy
  • Browser-based user interface

Product overview

Dedicated to live imaging, the Luxendo InVi SPIM is a microscope that has been optimized for long-term 3D fluorescence imaging of living specimens. Easy access to the sample chamber, maximized photo-efficiency, and short illumination times enable long-term imaging without harming live specimens.

The optical configuration combined with the fast acquisition speed of the InVi SPIM enables 3D reconstruction, tracking of cellular and subcellular positions, and morphological analysis in real time.

Typical applications of the microscope include in toto imaging of small animal models such as whole mouse embryos, Zebrafish and Drosophila embryos, imaging of dynamic processes in mammalian cell culture applications, and even live imaging of intact and living plant models.

Experimental excellence

The Luxendo incubation solution, as well as the sample chamber have been designed to provide close-to-natural environmental conditions for sensitive samples.

The incubation chamber provides temperature and humidity control as well as CO2 and O2 regulation. These ensure the long-term imaging stability for a wide range of samples of different origin.

The sample holder with its light-sheet-specific geometry is V-shaped and covered with foil that matches the refractive index of the immersion medium, i.e. water. As a result, an unobstructed projection of the light-sheet illumination and of observation view (path) is provided.

Specifications

  • Sample chamber
    • Water-sealed inert PEEK plastic chamber, autoclavable and biocompatible
    • Best-in-class low volume incubation
    • Easy access from above for sample mounting, injections, etc.
    • High-precision XYZ stage
  • Environmental control unit
    • Peltier-based water cooling and heating for precise temperature control
    • Temperature range: 20–37°C
    • CO2 range: 0–15 %
    • O2 range: 1–21 %
    • Humidity range: 20–99 % (gas humidifier with feedback control)

Device design

The InVi SPIM’s compact, vibration-free and robust design to provide 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.

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Illumination and Detection

The InVi SPIM can achieve a resolution down to 255 nm in xy, enabling subcellular resolution in living samples free of phototoxic effects.

The illumination optics comprise light-sheet generation by scanning a beam after passing a beam shaper unit for length and diameter adjustment. A Nikon CFI Plan Fluor 10x W 0.3 NA water immersion objective lens projects the light-sheet on the sample. The expandable and upgradable illumination setup allows a range of up to six lasers. The alignment-free light-source offers 40mW power output for each line.

The detection optics are optimized for fast, high-resolution and very sensitive imaging. A high numerical aperture Nikon CFI Apo 25x W 1.1 NA water immersion objective lens together with an additional magnification changer results in 31.3x and 62.5x total magnification for field view and sampling adjustment according to your experimental needs.

Illumination Detection Effective Magnification Field of View Pixel size Optical Resolution
10x/0.3 NA Nikon 25x/1.1 NA 31.3x 420 µm 208 nm 255 nm
    62.5x 210 µm 104 nm 255 nm

The two spectral detection channels equipped with 10 position high-speed filter wheels and a motorized dichroic mirror changer enable simultaneous dual channel imaging. In addition, more than two 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.

Compared to other techniques (e.g. 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 avoiding photodamaging effects, the downside of increased illumination in those techniques.

Specifications

  • Laser combiner
    • Maximum of 6 laser wavelengths (405, 445, 488, 515, 532, 561, 594, 642, and 685 nm with 40 mW each before fiber)
    • Fast modulation (MHz range) and high extinction ratio
  • Illumination optics
    • Chromatic correction from 440 to 660 nm
    • Light-sheet generation by beam scanning
    • Flexible light-sheet thickness (2 µm to 6 µm)
    • Nikon CFI Plan Fluor 10x W 0.3 NA water immersion objective lens
  • Detection optics
    • Nikon CFI Apo 25x W 1.1 NA water immersion objective lens
    • 2 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
    • 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

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 TCP/IP-based communication interface
    • 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

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.

Webinars

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Contact us

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