Dual Inverted Selective Plane Illumination Microscopy (diSPIM)

ASI has developed a new form of light sheet microscopy with our collaborators within the scientific community. The system utilizes two water emersion objectives mounted at 45 degree angles above the sample on an inverted microscope, a third objective also can be used to view the sample from below. The top objectives are mounted on piezo positioning devices so precise image stacks can be obtained. The system utilizes micro-mirrors to generate the light sheet, and the complete system can be mounted on to an existing inverted microscope, or supplied with ASI’s Rapid Automated Modular Microscope system (RAMM). Some of the benefits of the system include:

• Use conventional mounting /glass cover slips
• Generate 3D volumes with isotropic resolution (330 nm in all directions)
• Axial resolution is ~2x better than confocal- or spinning disk systems
• Achieve a ~7-10 fold reduction in photobleaching
• Acquisition rates up to 200 images per second or 2-5 volumes per second (roughly 10x faster than spinning-disk microscopes)
• Compared to Bessel beam plane illumination approaches, diSPIM offers equivalent (or better) axial resolution (~330 nm), 10-100x faster volumetric imaging rates (0.5-1 s instead of 10-100 s), and the ability to image over ~10x more time points (~1000 instead of ~100), due to the significantly lower illumination dose employed in our experiments and the lack of extraneous illumination outside the focal plane
• Microscope has multicolor capability, and has been tested successfully on cells cultured on cover slips, cells embedded in collagen gels, and nematode and zebrafish embryos.

For further details on ASI’s website: click here

For further information about diSPIM, check out the following link
https://www.ibiology.org/talks/selective-plane-illumination/

iBiology.com Microscopy Course about diSPIM presented by Dr Hari Shroff

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InVi-SPIM

Dedicated to live imaging- Looking downside up

Invi-SPIM system is dedicated to live imaging and is configured as an inverted microscope, 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 and cell cultures, and facilitates early embryonic development imaging. The unique optical configuration, in addition to ultra-fast acquisition speeds, enables 3D reconstruction, tracking of cellular and subcellular positions, and morphological analysis in real time. It utilizes light sheet technology, and includes an adjustable light sheet which is flexible in sheet area and variable in thickness 2-8 µm.

• Inverted microscope configuration
• Primary system for "in toto Imaging" of Mammalian Development from Zygote to Blastocyst.
• Easy access to Sample Chamber
• Maximized Photo-efficiency
• Long-term 3D Fluorescence Imaging of Live Specimens
• Close-to-Confocal Resolution
• Sample chamber and immersion medium separated
• Illumination objective: 10 x @ 0.3 NA, H2O immersion
• Detection objective: 25 x @ 1.1 NA, H2O immersion
• Small sample chamber volume
• Autoclavable, biologically inert sample chamber
• Accurate temperature control
• Optional environmental control (CO2, O2, N2)
• Easy sample accessibility for e.g. patch clamp, photoactivation/photoablation

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MuVi-SPIM

Life in the Fast Lane

MuVI-SPIM incorporates the latest sCMOS camera technology and optimized combinations of objective lenses to enable you to obtain the best from your sample. It provides four simultaneous orthogonal views on large living specimens, without the need for sample rotation, in order to dispel shadowing effects and facilitate long-term imaging at ultra-high acquisition speeds. The MuVi-SPIM is the fastest SPIM on the market for analysing dynamic processes, enabling 4 simultaneous views at 65 frames/sec. Ideal for applications such as developmental biology, embryonic development, marine biology, neurobiology, functional imaging, long time lapse imaging and large specimen.

• Large Field of View
• Close-to-confocal resolution
• Very High Imaging Speed;
• Great Sample Handling and Low Ototoxicity
• High Sensitivity and Low Noise
• 360 Illumination and Detection
• High-speed sCMOS camera Hamamatsu ORCA-Flash 4.0
• Laser combiner with six laser positions
• 2x Nikon CHI Plan Fluor 10XW 0.3NA water immersion for illumination
• 2x Nikon CFI LWD 25XW 1.1NA water immersion for detection
• Mounting Chamber with temperature control (15°C – 40°C)

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QuVi-SPIM: The Multi-View Light-Sheet Microscope

The QuVi SPIM is the latest product in Luxendo’s range of light-sheet microscopes. Combining dual views with dual detection channels, it enables large-scale imaging of living samples.Its novel stage design allows using SBS-format plates and provides an easy sample accessibility by means of its quick load feature. Since the objective lenses are exchangeable, a wide variety of samples of different sizes and preparations can be imaged.Typical applications of the microscope include living or fixed and cleared brain slices, long-term imaging of 3D cell culture models (spheroids, organoids, tumoroids), imaging of conventional cell culture in high throughput, and even functional (e.g. calcium) imaging.

Key Feautures

• Easy sample accessibility through quick load feature
• Compact, vibration-free and robust design
• Resolution down to 340 nm in 3D
• Browser-based user interface

Specifications

• 40x/0.8 NA Illumination
• Two Nikon 40x 0.8 NA water immersion objective lenses
• 40.0x Effective magnification
• 330 µm Field of View
• 160 nm Pixel size
• 340 nm Optical resolution
• 2 spectral detection channels, each equipped with a fast filter wheel (10 positions and 50 ms switching time between adjacent positions)
• 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
• Filters adapted to the selected laser lines
• Chromatic correction from 440 to 660 nm
• Flexible light-sheet thickness (2 µm to 6 µm)
• Maximum of 6 laser wavelengths (405, 445, 488, 515, 532, 561, 594, 642, and 685 nm with 40 mW each before fiber)

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