VIVO SDC

Upright Intravital Spinning Disk Confocal Microscopy System

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Upright Intravital Spinning Disk Confocal

The VIVO SDC platform incorporates advanced optics, cameras, fast excitation sources, computers and proprietary electronics to achieve high speed, precision and flexibility in intravital image acquisition. The configuration of the VIVO system is designed to allow the presentation of surgically prepared animals and live tissue slices without compromising high-sensitivity imaging at fast speeds. Seamless integration of ablation lasers and photoactivation/photoconversion devices allows for complex photomanipulation experiments.

Slide

Up to four fiber outputs
Millisecond path switching

Fully Automated Microscope

Platform Stage

Telecorr

Computer generated holography
1-photon and 2-photon

Intravital 3D confocal imaging
Super-resolution dual microlens disk

Motorized z-drive (with optional fast piezo)
Long working distance water dipping objectives
Large, easily accessible sample space

355nm and 532nm pulsed laser system
Fixed point or galvo-scanned for fast targeting

Accommodates intravital imaging trays and life support systems for long-term animal imaging

Modular high-speed X/Y scanner for accurate, diffraction-limited photomanipulation events

Millisecond timing and trigger
Control of multiple devices

Modular laser combiner
Up to eight lasers

Surgical Trays

Custom-designed surgical trays for presentation of exteriorised tissue and for stabilization of tissue presentation using cranial windows

Spinning Disk Confocal

Focus extender for correct spinning disk confocal excitation

Highlights

Complete Cardiovascular Imaging Platform

VIVO SDC is designed around fast imaging of the mouse vasculature for thrombosis and inflammation studies. Over the past two decades, dozens of high-impact journal articles have been published with the VIVO SDC system focusing on platelet and leukocyte biology in numerous murine vascular beds including dermis/skin-flap, mesentery, cremaster, cerebral-microvessels (via cranial window) and for imaging of larger vessels including the femoral and carotid arteries. 3i applications scientists can offer training and guidance from experimental design to publication based on extensive personal experience in animal preparation, surgical techniques, intravital imaging and data analysis.

Intravital Thrombosis Imaging

The Ablate! laser system is used to trigger gentle and reproducible damage to the endothelial cells in live blood vessels to induce platelet adhesion and thrombus formation. Consistent pulses from the laser allow for tissue ablation before and after addition of an agonist or inhibitor for detailed kinetic studies of novel therapeutic targets. Power can be easily titrated from cell activation to multi-cell ablations with a diffraction-limited spot for precise targeting.

Platelet thrombosis formation is at the center of many cardiovascular diseases, including heart attacks, strokes and deep vein thrombosis (DVT). VIVO SDC equips researchers with a reproducible laser-induced thrombosis model in vivo by implementing a sensitive camera, fast LED brightfield & fluorescence illumination, Ablate! pulsed laser and easy-to-use SlideBook software. In this experiment the murine cremaster muscle is visualized in brightfield and platelets are fluorescently labeled in red. After the ablation event a thrombus forms and stabilizes inside of the arteriole. SlideBook offers advanced masking, segmentation and statistical analysis that makes quantifying thrombosis data easy.

Surgical Equipment & Training Resources

3i offers extensive equipment lists and recommendations for experimental setups. Training videos and journal articles offer researchers a quick starting point for performing advanced cardiovascular imaging experiments. Surgical trays for tissue presentation with custom inserts are available for multiple animal models.

Forceps & Scissors

Superfusion

Dissection Stereomicroscopes

Thermostasis Equipment

Preclinical Research Pipeline

Intravital imaging has become essential in several fields including thrombosis & hemostasis, immunology, vascular biology, angiogenesis, biochemistry, gene therapy and cancer biology. Real-time imaging of cardiovascular events adds powerful in vivo results to studies of novel molecular mechanisms, accelerating drug discovery and decreasing the time to patient treatment. Intravital imaging with the VIVO SDC system is now being used to investigate and develop new therapies targeting hemophilia, leukemias and other cancers with circulating tumor cells, ischemic events, inflammation and other cardiovascular diseases.

 
THROMBOINFLAMMATION
The inflamed cremasteric venules of a WT mouse showing crawling neutrophils (white) capturing and releasing platelets (red) expressing CD62P (green). Courtesy of Dr. Vinatha Sreeramkumar & Dr. Andrés Hidalgo, CNIC.

Flexible and Robust Neuroscience System

Imaging the brain and nervous system of live organisms requires a gentle, fast and high-resolution system. VIVO SDC systems are used to image structure and calcium flux in live brain tissue slices as well as zebrafish and Drosophila brains. A large platform stage offers room for micromanipulators for neuronal patch clamp electrophysiology, paired with fluorescence imaging modalities through software triggering and synchronization capabilities. The addition of Phasor holographic photomanipulation enables rapid optogenetic stimulation while imaging.

Neurons in Spinal Cord Tissue
GFP expressing neurons in spinal cord tissue. Sample courtesy Dr. Wolfram Tetzlaff, University of British Columbia.

 
PTZ-induced seizures in the brain of Zebrafish larva
Calcium signals from the brain of Zebrafish larva in resting conditions and after PTZ-induced seizures. The transgenic line is Tg(NeuroD:GCaMP6f). Image courtesy of Laura Desban, Andrew Prendergast and Claire Wyart. Captured with 3i VIVO and Yokogawa CSU-X1 SDC.

Patch Clamp Electrophysiology

Patch clamp electrophysiology may be preferred for direct measurement of current and voltage changes taking place across the membrane in neural tissue. Several combinations of stages and micromanipulators can be paired with VIVO SDC systems including existing electrophysiology equipment.

Phasor Holographic Photomanipulation

  • Spatial light modulator-generated holography for optogenetics stimulation/FRAP/voltage imaging
  • Simultaneous 3D stimulation of multiple, separate regions
  • Visible and multiphoton stimulation without scanning
3D PHOTOSTIMULATION
3D illumination pattern (left) applied to a 3D specimen (right) to stimulate multiple regions simultaneously.

Increasing Imaging Depth with VIVO Multiphoton

An intravital imaging system for brain slice and in vivo multiphoton imaging, VIVO Multiphoton Upright is available as an upgrade to the VIVO SDC system. A flexible and modular design allows for the integration of best-in-class components from platform stages to scanheads to 2P holographic photomanipulation. VIVO Multiphoton is optimized for imaging deeper into smaller model systems (Drosophila, C. elegans, zebrafish, brain slices and organoids) as well as rodent animal models.

ZEBRAFISH
A maximum intensity projection of ciliated cerebrospinal fluid-contacting neurons (magenta) contacting the central canal in the spinal cord of a larval zebrafish. Cilia are labeled in green. Courtesy of Jenna Sternberg, Wyart lab, Institut du Cerveau et de la Moelle Épinière.

Technology

Spinning Disk Confocal

Yokogawa spinning disk confocals utilize a dual Nipkow disk with microlenses for the best optical sectioning and minimal pinhole crosstalk. This proven technology is the best solution for intravital imaging where optical sectioning and speed are both critically important.

CSU-X1

  • Highest speed imaging at up to 2000fps
  • Field of view for 7mm x 10mm detectors
  • 50µm pinhole disk with microlenses
  • Manual and motorized versions

CSU-W1

  • High speed imaging up to 200fps
  • Wide field of view 16mm x 17mm
  • 25µm and 50µm pinhole disks for lower and higher magnification objectives
  • Motorization including disk exchange, variable aperture, camera port selection and camera port magnification
  • Options for split-view imaging, NIR imaging, illumination field flattening and super-resolution imaging

CSU-W1 SoRa

CSU-W1 SoRa is an easy-to-use super-resolution microscopy solution utilizing a dual Nipkow disk assembly with microlenses on both the illuminating and pinhole disks. SoRa images have a 1.4x resolution improvement and deconvolved SoRa images have a 2x resolution improvement compared to standard spinning disk data. With a maximum speed of 200fps, low phototoxicity and no limitation on dyes or fluors, SoRa is ideal for super-resolution intravital imaging. SoRa is also available as an upgrade to existing CSU-W1 systems.

Imaging of microtubules in fixed bovine pulmonary artery endothelial cells. Azuma, T. and Kei, T. (2015) Super-resolution spinning-disk confocal microscopy using optical photon reassignment. Opt Express. Jun 1;23(11):15003-11. doi: 10.1364/OE.23.015003.

SlideBook Software for Acquisition and Analysis

SlideBook software supports research microscopy through the entire experimental process. By managing everything from instrument control to image processing and data analysis, SlideBook allows scientists to focus on investigation rather than instrumentation. SlideBook controls hundreds of instruments in and around the microscope from dozens of manufacturers enabling researchers to integrate their preferred components and upgrade to the latest devices once available.

Slide

User-Selectable App Appearance

Select a color scheme from dozens of options
Switch on-the-fly from dark to light themes

SlideBook Open File Format

Directory-based open file format for big data and high performance computing applications

Volume Rendering

3D and 4D volume view visualization tools support a user- specified bounding box and a storyboard interface where multiple perspectives can be assembled into a single movie

NVIDIA CUDA GPU Acceleration

GPU acceleration of computationally-intensive operations such as deconvolution

VAST Integration

SlideBook communicates with the VAST BioImager platform for automated micron-level positioning of large specimens

3D Capture Status

Montage

Volumetric projection during 4D capture supported across all instruments

Acquire and stitch 3D volumes across multiple fields of view

Capabilities

Capture

Control hundreds of devices including microscopes, stages, lasers, wheels, piezos, scanners, shutters and much more.

View

Visualize data through any numbers of portals, from single images to z-stacks, time lapse, color channels and 4D views.

Analyze

Analyze images and extract statistical data via a wide variety of algorithms while maintaining original data integrity.

Scripting

Macro scripting for capture and analysis enhances the flexibility and power available to users.

Communicate

Present and export data easily as 16-bit TIFFs, 3D movies, graphs or spreadsheets. Data is directly portable to MATLAB and Excel and adheres to Open Microscopy Environment (OME) standards.

Partners

MATLAB

Through hierarchical and conditional capture, user-supplied MATLAB programs can control experimental workflows.

Microvolution

Microvolution software delivers nearly instantaneous deconvolution by combining intelligent software programming with the power of a GPU.

Aivia

Aivia is an innovative and complete 2D-to-5D image visualization, analysis and interpretation platform with artificial intelligence-guided image analysis.

Dell

The latest high-power computer workstations control all microscope hardware and enable high-speed processing, segmentation and volume rendering of terabyte (TB) datasets.

Powerful Computer Workstation

3i provides high-power computer workstations to control all microscope hardware necessary for acquisition as well as enable processing, segmentation and volume rendering of terabyte (TB) datasets without additional computer resources.  Solid state drives in RAID configurations provide high-speed storage for capture, while traditional high-capacity hard drives provide longer-term storage of datasets. NVIDIA Quadro GPUs work in parallel with the latest Intel Xeon processors for fast stitching, processing and rendering of captured data.

Petabyte Data Storage

3i offers DDN® unified storage systems to allow direct acquisition and analysis, without time-consuming file transfers, at volumes ranging from 500TB to over 2PB. DDN storage systems are an ideal choice for labs and facilities looking to optimize acquisition workflows and/or incorporate data analysis pipelines.

VIVO SDC Data Sets

 
PULMONARY INFLAMMATION
Large vascular beds, as seen in the murine lung, allow for macroscale imaging of leukocyte adhesion, accumulation, and transmigration in real time. Mouse lung vasculature (CD31, blue) and leukocytes (yellow) imaged with a VIVO system. Image courtesy of Dr. Dean Kavanagh, ICVS, Birmingham University.
 
thrombosis
High-speed 3D imaging is required for visualizing thrombosis formation in vivo. Platelets (red) and fibrin (cyan) imaged after laser-induced thrombus formation Image courtesy Dr. Vivien Chen, University of Sydney.
 
Zebrafish Vascular Network
Small model organisms are a great substitute for mouse imaging while still offering insight into the inflammatory response. Zebrafish vascular system (green) and leukocytes (red) imaged with a VIVO SDC.
CARDIAC CIRCULATION
Murine heart vasculature visualized with FITC-BSA (green) and leukocytes with anti-Gr-1 antibodies (red). Image courtesy Dr. Dean Kavanagh, University of Birmingham.
Thrombosis
Mouse cremaster arteriole after laser-induced thrombus formation. Vasculature (green) and platelets (red).
 
TRANSMIGRATION AND EXTRAVASATION
The inflammatory response begins with neutrophil recruitment to the site of infection and/or injury. Leukocytes (red) after leaving cremaster microvasculature following perfusion of chemoattractant.
 
Cell segmentation and tracking
Neutrophil rolling and crawling inside of mouse vasculature can be easily labeled and quantified using the robust particle tracking tools in SlideBook.

Additional Resources

Brochures

VIVO SDC

Webinars

 

VIVO Intravital Microscopy Platform for Cardiovascular Studies

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    SALES

    Email: sales@intelligent-imaging.com

    Phone: +1 (303)-607-9429 x1

    SUPPORT

    Email: support@intelligent-imaging.com

    Phone: +1 (303)-607-9429 x2