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Andor Revolution XD brochure

Andor Revolution XD brochure

Product catalog summary

Introduction
The document introduces the Revolution series of confocal spinning disk solutions by Andor, emphasizing their high-speed imaging capabilities and suitability for live cell imaging. The systems are built around Yokogawa's CSU spinning disk technology, known for its sensitivity and low phototoxicity.

Key Technologies
The Andor iXon ultra and sCMOS cameras are highlighted for their high-speed and high-resolution capabilities, essential for capturing detailed cellular dynamics. The systems offer flexibility with various configurations to meet specific research needs.

Revolution WD and XD Systems
The Revolution WD system is designed for versatility, offering a large field of view and optimized performance for thick samples. It is suitable for a wide range of applications, including developmental biology and neuroscience. The Revolution XD system focuses on high-speed, high-sensitivity imaging, ideal for applications like ion imaging and vesicle tracking.

System Configurations
The document details various configurations within the Revolution series:

Revolution WDi: A versatile solution for inverted microscopes.
Revolution XDi: A high-speed configuration for inverted microscopes.
Revolution XDh: Combines XD system with an upright microscope.
Revolution XDv: A compact solution for upright microscopes, suitable for labs with limited space.

Active Illumination and Applications
The active illumination portfolio includes tools for research in areas like cell signaling and optogenetics, using techniques such as photo-bleaching and uncaging. The systems are designed to meet the needs of various research fields, including cancer research and plant biology.

Features and Benefits
The document lists features such as the Yokogawa CSU spinning disk unit, iXon EMCCDs, and sCMOS cameras, highlighting benefits like high speed, low phototoxicity, and large fields of view. Additional tools like the tuCam dual camera adapter and Andor's laser combiner enhance the systems' capabilities.

Custom Solutions
Andor offers bespoke services to meet specific customer requirements, providing custom mechanical and optical designs. This flexibility allows integration with existing hardware for cost efficiency.

Conclusion
The Revolution series by Andor provides advanced solutions for high-speed, high-sensitivity live cell imaging, with customizable options to suit diverse research needs.

Camera Technologies

EMCCD Cameras: Provide high sensitivity for low light conditions, minimizing phototoxicity and detecting weak signals.
sCMOS Cameras: Offer high-speed imaging with low read noise, suitable for detailed confocal resolution.
Interline CCD: Known for higher noise levels compared to sCMOS and EMCCD.

Software Solutions
The document outlines various software options for data acquisition and analysis, including:

Imaris: For visualization and interpretation of multidimensional datasets.
Andor iQ: Offers flexible protocol architecture and Python scripting for complex experiments.
MetaMorph: Supports a wide range of third-party components.
MicroManager: An open-source solution for hardware control and image capture.
NIS Elements: Provides a comprehensive GUI for Nikon microscopes.
LabVIEW: Allows custom software development for specific hardware needs.

Microscopy System Components
The document details various components and their features:

Confocal Spinning Disk Units: CSU-X1 and CSU-W1 offer high-speed imaging with low phototoxicity.
Filter Wheels: Allow wavelength or polarization selection with high-speed switching.
FRAPPA: Provides fluorescence recovery and photoactivation capabilities.
MicroPoint: Offers laser illumination for ablation and activation.
Stage Incubator: Maintains suitable environments for cell cultures on the microscope stage.
Laser Combiner: Supports multiple laser lines for confocal imaging and other applications.
Mosaic Active Illumination: Utilizes DMD technology for precise illumination control.

Dual Camera Solutions
The TuCam dual camera adapter allows simultaneous dual-color imaging and easy switching between cameras with different attributes.

Applications and Technical Notes
The document includes examples of research projects utilizing spinning disk confocal systems, emphasizing their strengths in live cell imaging experiments.

Specifications and Procedures
The study utilized Drosophila S2 cells expressing CID–mCherry and GFP–α-tubulin to visualize centromeres and microtubules. Confocal fluorescence microscopy was employed, but visualization was challenging due to the small size and low fluorescence of kinetochores. The Andor Revolution XD spinning disk confocal microscopy was used to address these challenges, focusing on speed of acquisition, spatial and temporal resolution, and fluorescence intensity management.

Key Findings
In the absence of POLO kinase, Drosophila cells exhibited syntelic chromosome attachment, indicating a lack of corrective mechanisms for proper chromosome alignment. The presence of POLO kinase was shown to ensure correct centromere architecture and chromosome bi-orientation.

Technological Insights
The spinning disk confocal microscopy provided superior spatial resolution and allowed real-time high-contrast imaging at the diffraction limits of the microscope's optics. This enabled detailed 3D visualization of kinetochore behavior and spindle attachments.

Applications in Gene Territory Mapping
Dr. Gadal's research on yeast cells used spinning disk confocal microscopy to map gene territories in a 3D format, overcoming limitations of fixed cell observations. The study highlighted the importance of gene positioning within the nucleus for gene expression and regulation.

Conclusion
The use of spinning disk technology in live cell applications revealed critical insights into chromosome arrangements and gene territories, emphasizing the importance of nuclear architecture in genomic functions.

Confocal Microscopy Techniques
1. Confocal Laser Scanning Microscopy (CLSM): Utilizes a laser to scan the sample point by point, offering high resolution but with a trade-off between speed and image quality.
2. Spinning Disk Confocal Laser Microscopy (SDCLM): Overcomes CLSM limitations by using multiple pinholes to illuminate the sample simultaneously, enhancing sensitivity and speed.

Technical Advancements
The document details the dual spinning disk confocal laser scanner, which uses microlenses and pinholes to improve excitation throughput and reduce background noise. The use of high quantum efficiency detectors like EMCCD cameras further enhances performance.

Fluorescence Anisotropy
This technique measures molecular interactions and dynamics within cells. The document introduces a spinning disk confocal anisotropy system (SpiDA) that allows high-definition anisotropy imaging by overcoming polarization degradation issues in standard confocal systems.

Applications and Limitations
The document discusses the use of anisotropy imaging in studying molecular organization and interactions, particularly in living cells. It also addresses the challenges of maintaining polarization fidelity in confocal systems and presents solutions to enhance imaging capabilities.

Conclusion
Confocal microscopy, particularly with advancements like SDCLM and SpiDA, provides powerful tools for detailed biological imaging, offering insights into cellular processes and molecular interactions.

Technical Specifications
The document highlights the use of a dual-camera adapter, TuCam, which ensures alignment stability and low distortion. The system employs high-performance EMCCD cameras for high signal-to-noise anisotropy imaging, crucial for studying protein interactions and microstructure modulation.

System Components
Key components include a motorized polarizer, a high-quality polarizing beam splitter, and a kinematic assembly for precision and stability. The system's mechanical stability is critical for accurate and repeatable measurements.

Imaging Performance
The SpiDA system reduces out-of-focus fluorescence, enhancing resolution and dynamic range. It uses back-illuminated sensors with a significant cost premium, making multi-megapixel expansion costly.

sCMOS Technology
sCMOS technology offers high fidelity and quantitative measurement capabilities, overcoming limitations of traditional CCDs and EMCCDs. It provides a large field of view, high resolution, and low read noise, making it suitable for high-speed, low-light applications.

Comparison with Other Technologies
The document compares sCMOS with Interline CCD and EMCCD technologies, highlighting sCMOS's ability to maintain low noise at high frame rates. EMCCD technology, while sensitive, introduces multiplicative noise, affecting signal-to-noise ratio.

Conclusion
The Andor solution for real-time confocal anisotropy imaging addresses constraints in polarized excitation of fluorescence. The SpiDA system, with its dual-camera adapter and sensitive cameras, provides high-contrast dynamic anisotropy data for live cell studies.

Acknowledgments
The document acknowledges Dr. Satyajit Mayor and his team for their support and collaboration.

References
A list of references is provided, supporting the technical content discussed in the document.

Specifications and Comparisons
sCMOS technology offers a high dynamic range of 30,000:1 at 30 fps, which is superior to Interline CCDs and EMCCDs. The architecture of sCMOS allows for a large well depth despite small pixel sizes. A comparative overview in Table 2 shows sCMOS outperforms Interline CCD and EMCCD in most parameters, including read noise, frame rate, and quantum efficiency.

Performance Analysis
Figures 1 to 8 illustrate the performance of sCMOS compared to Interline CCD and EMCCD under various conditions. sCMOS demonstrates better signal-to-noise ratio (SNR) and resolution, especially in low-light conditions. Theoretical SNR plots further support the superior performance of sCMOS over Interline CCD across a range of photon fluxes.

Conclusion
sCMOS technology represents a significant advancement in scientific imaging, offering low noise, fast frame rates, wide dynamic range, and high resolution. While EMCCD remains advantageous for extremely low-light applications, sCMOS is poised to gain widespread recognition for its overall performance.

Customer Support and Maintenance
Andor provides extensive customer support services, including on-site assistance, training, and testing services. Various maintenance contracts, such as Platinum and Silver, offer priority service, remote support, and preventative maintenance visits to ensure optimal product performance.

Catalog excerpts

Confocal Spinning Disk Solutions Setting the standard in high speed imaging

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Meet the family Experience tells us that almost every customer is unique in their requirements, and so rarely are two Revolution systems the same. We pride ourselves in offering flexibility so that you can focus on your applications. The Revolution WD sets new standards in spinning disk confocal microscopy, and brings this renowned live cell technology right up to date for live cell studies. With the large field of view and improved performance with thick samples, the WD brings outstanding versatility for a broad range of applications. The Revolution XD is the ultimate solution for high speed,...

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The Revolution WD is the latest spinning disk confocal system in Andor’s range. It is the ultimate solution for the diverse range of samples used in live cell imaging. Using the CSU-W1 from Yokogawa, the Revolution WD retains the pedigree expected from the Revolution systems, but offers so much more in order to meet your research needs. TuCam CSU OptoSplit Key Applications Regenerating cholinergic motorneurons in the nematode C.elegans. Upper Image - Dual labelled nematode. Collaboration with T. Edwards, Hammarlund Laboratory, Yale School of Medicine. Camera Piezo Stage Developmental biology...

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The Revolution XDi is based around an inverted microscope, the most common choice for live cell imaging. With a disk speed that supports up to 2,000 frames per second, it is the primary choice for high speed cellular dynamics and applications such as ion imaging. It delivers premium performance for speed and sensitivity at high magnifications. Flexibility The Revolution XD is more than just a spinning disk solution. For membrane related biology we can integrate with TIRF illuminators. Active illumination devices offer a deeper understanding of cell dynamics and signalling. Spinning disk technology...

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Full Featured Upright Microscope System XDh has been designed to allow those who work on an upright microscope to benefit from the capabilities available from our full component range. The XDh is suitable for applications such as plant biology research, and in developmental biology models such as c.elegans and drosophila. In these cases, whilst an inverted microscope configuration is possible, an upright microscope can be more practical owing to the sample preparation with a glass coverslip more typically on the upper facing surface. Other examples of applications requiring an upright microscope...

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Revolution XDv Upright Scope with Reduced System Footprint The Revolution XDv offers you a compact solution on an upright microscope. In keeping this system footprint small, it is well suited to labs with limited space or to fit inside a Faraday cage for electrophysiology rigs. In securing the CSU-X, filter wheel and camera to our vertical mount, you are guaranteed complete stability. Filter Wheel Laser 3 Laser 4 Laser 5 Laser 6 Key Application Example - ELECTROPHYSIOLOGY With the Revolution XDv you can confidently combine spinning disk confocal microscopy with techniques as sensitive as patch...

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Revolution Custom Systems The flexibility for me to define a novel configuration for the Revolution XD allowed me to combine live cell imaging with laser tweezers. This offers new avenues of investigation for my research. At Andor we realize that, sometimes, even our adaptable and flexible off-the-shelf products are not enough to meet some of the more demanding application requirements of our customers. For this reason we provide a bespoke service to our customers, whereby a dedicated highly experienced team of engineers and application specialists provide customer specific solutions. The process...

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Microscopy System Cameras EMCCD technology revolutionized spinning disk confocal microscopy. The ability to cool to -100°C and the use of EM gain effectively eliminates noise and provides routine single photon sensitivity. The key benefits of Andor’s iXon EMCCD cameras are: The Neo sCMOS camera is the perfect partner to the iXon range. Though not as sensitive as EMCCD cameras, sCMOS runs at high speeds and has extremely low read noise at 1 e-/pix/sec, giving excellent signal to noise performance. The key benefits of Neo sCMOS are: • Extreme sensitivity for working at low levels of sample illumination,...

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Software Flexibility in your choice of software as well as hardware. Instrument Control and Data Acquisition Data Visualization and Analysis Bitplane Imaris iQ’s flexible protocol architecture can be easily tailored to your specific experiment, including ratio-metric imaging, FRAP analysis and other complex setups. Its unique image disk can handle massive 6D data sets. Molecular Devices’ MetaMorph software is a popular and familiar interface for many users. Metamorph supports a broad range of third party components including the Revolution WD/XD and Active Illumination portfolio. Imaris delivers...

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Microscopy System Components Confocal Spinning Disk Unit Confocal Spinning Disk Unit The CSU-W1 offers complete versatility, addressing a wide range of sample types due to an alternative disk design to the CSU-X1. Choose up to 2 disk patterns according to your requirements. Available in 3 models: single and dual camera, and split-view, the CSU-W1 offers a range of options according to your budget or application requirements (e.g. simultaneous dual wavelength capture). Upgrade single camera versions in the future with TuCam if you wish. (see page 23) The CSU-X1 from Yokogawa is the premier solution...

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Microscopy System Components Rotr™ Filter Wheel - Emission discrimination Mosaic 3 - Active Illumination Our systems use filter wheels to select by wavelength or polarisation. Our new highspeed Rotr filter wheel offers wavelength switching up to nine frame pairs/sec when fully loaded with filters (manufacturers quote unloaded times). Multiple filter wheels are sequential. The Mosaic active illumination system utilizes digital mirror device (DMD) technology to control the illumination field of a fluorescence microscope. Using laser or arc lamp sources, Mosaic achieves real time and near diffraction...

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Andor Revolution XD brochure

Andor Revolution XD brochure

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