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VIVO™ is the ideal turnkey imaging system for live-animal intravital imaging. VIVO™ facilitates various quantitative microscopy techniques for the measurement of dynamic processes in a light-starved environment. Both practical and easy to use, VIVO™ is designed for speed and sensitivity to visualize metabolic processes in live cells. VIVO™ is compatible with multiple imaging modalities including transmitted light, fluorescence and confocal imaging on a single microscope with minimal manipulation.

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Applications

Common uses of VIVO™ include real-time multi-dimensional imaging (6D applications), ratiometric imaging, FRET, 3D deconvolution, timelapse, and more in a low light environment. Advanced options further expand the capabilities of this system give you the ability to address a wide variety of applications. Some of the advanced options available include: spinning disk confocal, environmental control, FRAP, FLIP, photoactivation and uncaging, TIRF, FLI, single molecule studies, real-time deconvolution, rapid 4D, simultaneous dual camera acquisition and synchronization with electrophsiology and automated perfusion systems.

Intravital imaging (imaging within a living animal) has become realized in several fields including Thrombosis and Hemostasis, Immunology, Vascular Biology, Angiogenesis, Biochemistry, Gene Therapy, Development, and Cancer Biology.

• The imaging of blood clot formation and the recruitment and involvement of blood coagulation proteins was first captured in real time in multiple channels on a VIVO™ system.
• The 3D reconstruction over time of the forming blood clot was captured with multiple fluorescent probes using VIVO™, both widefield deconvolution and Spinning Disk Confocal.
• VIVO™ used with Laser Ablation, Ferric Chloride, Rose Bengal (photo) Injuries.
• VIVO™ with particle tracking has been used to capture and count white cell rolling along a vascular tree.
• VIVO™ is assisting in the formation of new targeted therapies for Hemophilia and other blood related genetic diseases.
• The identification of cancer tumor markers and their roles in physiological processes have been recorded using VIVO™.
• Blood vessel sprouting can be recorded over time and computed with VIVO(tm) and Slidebook software.
• Enzymologic parameters using fluorescent FRET substrates introduced into tissues of living animals are imaged with VIVO™.
• Calcium transients in circulating cells have been measured using VIVO™.
• Intracellular Signal Transduction phenotyping in vivo
• Pharmaceutical Therapeutic Targeting and Screening in live animals

Organisms that have been imaged with VIVO™ include Bacteria, Yeast, Drosophila, Zebrafish, various Embryos, Mice and Rats. Mammalians have been imaged in the Carotid Artery, Cremaster Muscle, Messentary Muscle, Ear, Dermis, Eye, and Brain to date.

Complete System Motorization

VIVO™ is designed to have complete computer control of the entire microscope. This allows our SlideBook™ software to easily orchestrate complicated data collection protocols involving illumination control, filter selection, position regulation, detector selection, etc. The points of computer control and the precision of automation are listed below:

• X,Y-Axis (100 nm reproducibility due to linear encoders)
• Z-Axis (25 nm minimum step, 50 nm reproducibility due to harmonic drive)
• Fluorescence shutter (2 ms timing)
• Fluorescence intensity (2 ms timing, 8 settings)
• Fluorescence Excitation filter (4 positions, 2 ms switch time)
• Fluorescence/Reflected light filter turret (5 positions, 0.5 second switch time)
• Objective turret (automatic parafocal offset)
• Transmitted light turret (bright field, dark field, DIC and phase ring position)
• Transmitted field diaphragm
• Transmitted aperature diaphragm
• Transmitted light intensity
• ND filter
• Camera position prism
• Spherical Aberration Correction Setting

System Components

• Fixed-stage upright microscope
• DG4 - 300W upgrade recommended
• Micropoint Photoactivation, Photobleaching, Tissue Ablation System - galvanometer driven option recommended
• Uniblitz High Speed Brightfield Shutter
• High-speed digital camera
• Videoscope Image Intensifier
• Yokogawa CSU-X
• Roper QuantEM SC
• Multi-line AOTF controlled LaserStack laser launch¹
• Slidebook Software
• Latest Dell workstation and Nvidia graphics

Options

• Rapid 4D option - Peizo-electric focusing collar
• MultiCamera Option - DualCam + second QuantEM SC or 9300-201
• Simultaneous 2 color capture - DualView image splitter

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Critical Fluorescence Illumination - VIVO™ is equipped with a scrambled and stabilized 175 Watt Xenon light source designed to fill the back aperature of objectives delivering an intense, uniform and stably illuminated field. The light scrambling is particularly important for deconvolution applications because it makes illumination axially and laterally uniform. This keeps the Point Spread Function (PSF) the same over the imaged volume.

Spherical Aberation Correction - Spherical Aberration is the most serious monochromatic aberration in microscopy. It arises when the observed sample does not have the exact cover slip, mounting medium and immersion medium specified by the lens designer. Practically, this occurs with every biological sample.

VIVO™ comes standard with a patented motorized spherical aberration system from Infinity Photo-optical. This system, exclusively available from 3i, along with SlideBook™ for VIVO™ automatically adjusts optics to dynamically correct for spherical aberration for every focal plane in the sample. This yields data with the highest possible resolution and greatest contrast. These aberration free data are optimal for Deconvolution and 3D morphometrics.

Custom Optimized Optics Selection - Every scientific experiment is different and has a different set of optical requirements for optimal data collection. Each VIVO™ system comes with optics specifically tailored to each researcher's individual requirements.
System considerations include magnification, wavelength transmittance, working distance, DIC or phase optics requirements, and fluorescence probe requirements. Each VIVO™ system includes optimized filter sets that are thickness and wedge matched to have no lateral chromatic shift. In addition, filters are anti-reflective coated to maximize transmittance and optimize optical performance.

State-of-the-art Detectors - The camera on our VIVO™ system is optimized for sensitivity and collection speed. The sensor is a Sony extended quantum efficiency interline SVGA CCD. The detector is cooled to reduce thermal noise. The cooling fan has been externalized to eliminate image distortion via vibration.
VIVO™ can be equipped with additional detectors such as Cascade cameras or ICCD's for certain applications like rapid 4D collection. A second standard camera can be added to VIVO™ for simultaneous dual emission collection for experiments such as live FRET.

SlideBook™ for VIVO™ - At the heart of every VIVO™ system is our state-of-the-art SlideBook™ software. SlideBook™ for VIVO™ includes all the high-end functionality to fully take advantage of the system's automation. SlideBook™ seamlessly integrates x-,y- and z- stage motion with filter control and light shuttering to easily configure experiments as simple as 2D capture to as complicated as 4D multifluor and multi-mode imaging.

1. Complies with FDA performance standards for laser products except for deviations pursuant to Laser Notice No. 50, dated July 26, 2001