High-content Screening Station for Life Science
ScanR is a modular microscope-based imaging platform designed for fully automated image acquisition and data analysis of biological samples. ScanR can handle many different formats, including multi-well plates, slides and custom-built arrays. This unmatched flexibility makes it equally adept at routine and advanced applications, while preconfigured assays for common applications improve efficiency. Its powerful analysis module provides complex image analysis and advanced data evaluation for standard and bespoke assays.
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For some challenges, hard work and motivation alone are not enough, and the ideal equipment is essential. The scanR screening station is the perfect solution to the challenge of microscope-based high-content screening. scanR perfectly combines the modularity and flexibility of a microscope-based setup with the automation, speed, throughput, reliability, and reproducibility demands of screening applications. The system is thus equally well suited to handling standard assays and assay
development. Furthermore, the modular design allows the system to be adapted to the specific applications of an R&D lab as easily as to those of multiuser environments.
Designed in collaboration with the EMBL (Heidelberg, Germany), scanR was developed as a powerful screening station with an extremely broad application range complemented by sophisticated image- and data-analysis software, which is based on a powerful, interactive, cytometry-oriented approach for handling and analyzing huge numbers of multidimensional data sets.
The Olympus ScanR Screening Station for Life Sciences research combines the modularity and flexibility of a microscope-based setup with the automation, throughput, reliability and reproducibility required for screening applications. The system is suited to handling standard assays and assay development workflows. Furthermore, the modular design allows the system to be adapted to the specific applications of an R&D laboratory as easily as to those of multi-user environments. ScanR has been developed as a powerful screening station with a broad application range on the basis of experimental screening systems designed by the European Molecular Biology Laboratory (EMBL).
Designed for fully automated image acquisition and data analysis of biological samples, ScanR can handle many different formats, for example, multi-well plates, slides or custom-built arrays. The unmatched flexibility and open design make it adept at both routine and advanced applications. With a powerful analysis module for biological functional assays, it is an ideal tool for assay development and high-content screening. The ScanR provides complex image analysis and advanced data evaluation abilities, enabling it to address a whole range of standard and bespoke assays.
The system handles fixed and live cells with equal ease and is therefore the perfect screening platform for a wide cross section of research. ScanR specifically targets the need for quantitative imaging and image analysis in modern cell biology, systems biology and medical research. For example, cancer research involves investigations on many levels, such as DNA damage, cell proliferation and cell cycle analysis. Furthermore, novel genome-wide screens using RNA interference are a key
application to which ScanR is especially adapted (developed in close cooperation with the
EMBL). For drug screening, a great many protocols have been developed to show the biochemical effects of compounds on the cellular level. For example, drug-induced changes in gene expression levels can be assessed accurately via a range of standardized assays. ScanR also excels at routine screens measuring apoptosis, micro nuclei or DNA fragmentation (comet assays). Additionally, ScanR covers a huge range of screening applications (selected examples are listed below) which are implemented in many research areas.
Each cell-based experiment produces a plethora of information, most of which is not fully collected because many assays are completed as end-point studies with fixed cells. What is missed, though, are the dynamic cellular and sub-cellular events that are key to understanding the causes or effects on the molecular level.
ScanR provides time-lapse screening functionality to track the dynamic changes over time in tens of thousands of living cells.
ScanR incorporates advanced modular software and hardware, controlled through an easy-to-use interface that provides access to all image acquisition and image analysis parameters. The system includes a unique real-time controller which synchronizes the ScanR hardware and the unique multifunctional Olympus MT20 illumination system. The combination of these innovative devices ensures maximized acquisition speed, minimizes bleaching as well as phototoxicity, and provides the basis for true quantitative measurements. The system is designed to produce multidimensional (X, Y, Z, t, λ) images with great ease. Once acquired, images can be automatically analyzed by the powerful ScanR image and data analysis software that is based on a cytometry-oriented approach towards the handling and analysis of huge amounts of multi-dimensional data sets. The ScanR analysis software allows for full image processing (e.g. background correction), object and sub-object detection, parameter calculation as well as sophisticated gating and classification schemes.
To extend the functionality of ScanR, a series of optional components can be added. For example, the Olympus cellVivo environmental control system enables long-term live cell imaging. An optional plate-loading robot greatly enhances automation. Other optional features such as an IR laser-based autofocus, LED-based fluorescence illumination and significantly extended field of view add even more flexibility. Additionally, ScanR enables integration of customized and customer-built analysis modules for researchers in need of particular analysis capabilities.
The Olympus ScanR software features an intuitive graphical user interface based on a strict, workflow-oriented approach. This ensures simple handling in daily operation, easy image acquisition and straightforward system configuration. The ScanR system design is focused specifically on quantitative measurements and flexibility, addressing the needs of scientific screening and assay development.
scanR is based on the latest Olympus high-end IX83 inverted microscope, providing the utmost flexibility, robustness, and stability. It handles all standard assay formats, such as micro well plates and slides, with ease and can also be configured to accept any custom designs, such as spotted arrays or biochips.
Highly Stabilized Illumination
The MT20 illumination system developed by Olympus has, at its core, a highly stabilized xenon or mercury/xenon burner, a fast filter wheel, an achromatic and absolutely reproducible attenuator, and a fast shutter with an exceptional on/off time of less than 1 ms. Optionally, scanR can be operated with a high-end LED illumination system for maximal switching speed and long-term intensity stability.
The extreme demands for robustness, reliability, and throughput as well as minimized photobleaching are met by the perfect device synchronization and parallelization of the unique real-time controller (U-RTC) from Olympus.
The advanced features of scanR allow truly multidimensional (X, Y, Z, t, l) screening. Time-lapse Z-stack images can be recorded at any number of locations on micro well plates, slides, or custom formats using all available observation methods, including fluorescence or brightfield, as well as differential interference contrast (DIC) or phase contrast.
Fast and accurate autofocus is crucial for successful automated image acquisition. scanR provides a combination of hardware-based autofocus together with different software autofocus algorithms to meet the demands of extremely variable biological samples.
Large Field of View
Equipped with a large sensor-area camera and an optimized fluorescence condenser, scanR can acquire huge areas of the sample with each camera image, increasing the amount of acquired data and decreasing the screening time dramatically.
Based on an initial pre-scan, the scanR Analysis software can determine all interesting objects with the complete image processing, image analysis, and cytometric data exploration engine available. In an automated workflow, the analysis results are employed to scan selectively only the objects-of-interest in a second, targeted screen. Typical scenarios where Multi-level Acquisition excels are large area samples with few cells which require high resolution or single cell events.
With the huge amount of data that can now be collected from assays comes the need for coherent and careful quantitative automated analysis. The powerful ScanR analysis software package is completely independent from the ScanR acquisition software. This enables both modules to be installed on either the same or different workstations, connected via a local network. In both cases, image and data analysis can be performed “online” at the same time as acquisition or “offline” on previously acquired data sets. By collecting more data, the number of analytic techniques that can be carried out on samples can be increased. These can be as simple as counting the labeled cells on display or as complex as ratiometric feature-based analysis of multilabeled objects and sub-objects in different cell types. Image analysis is carried out as a logical multi-step procedure consisting of image processing, object detection, feature extraction and data analysis by gating and classification. A workflow-oriented user interface guides users through all steps.
Before nuclei, cytoplasm, and other subcellular objects are contoured, the raw images are preprocessed if necessary. For example, Adaptive Size Background Correction or calibration-based Shading Correction are used to remove heterogeneous background and shading automatically and rigorously while retaining the relevant intensity information. Spectral Unmixing can remove potential bleed through of different fluorophores effectively.
Object Detection and Analysis
Powerful object detection modules are optimized to segment nuclei, cells or other structures. Several detection algorithms can be selected and adapted to the objects of interest. Based on this, segmentation-relevant features can be selected out of a list of more than 100 object parameters. The selected parameters are extracted for evaluation and classification. Additional mathematical operations can then be performed on the parameters. This highly flexible data output enables the ScanR system to be used for almost all current and future cell-based assays.
Gating and Classification
ScanR specifically excels in subsequent data analysis and evaluation. For this purpose, the powerful data analysis concepts that are successfully applied in cytometry are adapted to the specific demands of analyzing large image data sets. The multidimensional image data generated are displayed in two-dimensional scatter plots or one-dimensional histograms. From these, clustered data populations of interest can be selected via graphical tools. Selected data sets can be repeatedly gated for further investigation. Gated objects can be re-scanned to perform automated rare-event analyses, for example.
Immediate Quality Control
Images and objects are linked reciprocally to any data point related to them. By clicking on a data point, the respective image is loaded into the display window and the respective object is highlighted. By clicking on an object in the image display window, the related data points in the scatter plots and histograms are highlighted. Additionally, a gallery view of all images of a selected or gated data population can be created to allow a direct and visual comparison of larger image sets with relevant information.
scanR was designed not only to satisfy the specific speed, endurance, and reliability requirements of a fully automated high-content screening system but also to provide unmatched flexibility and adaptability coupled with extensive expansion capabilities. This enables scanR to perfectly match the specifications of any application and budget.
Time-lapse Cytometry – Measuring Kinetic Parameters
The scanR kinetic module allows objects to be classified by their kinetic properties. Tracking curves are evaluated on the basis of values (mean static parameters such as intensity, area, ratio, shape factor, etc.) measured over time. The curves are condensed into single characteristic values. Finally, kinetic parameters can be plotted in 1D or 2D histograms, and populations can be gated based on their specific kinetic properties.
The 3D Deconvolution module is a powerful image processor which uses known optical parameters (e.g. numerical aperture, wavelength) to optimize image quality, remove out-of-focus light and improve optical sectioning.
A Tale of Two Systems
scanR is compatible with the Olympus cellSens live cell imaging software, which can run on the same system. The same setup can thus be used both as the dedicated scanR screening system and at the same time as a high-end imaging system.
Plate-loading Robot and Integration Interface
The scanR platform supports a plate-loading robot. The robot facilitates fully automated screening of multi well plate stacks and extends screening to 24/7 operation. scanR offers flexible support for integration into automated sample preparation lines.
IR Laser Hardware Autofocus based on IX83 ZDC
The latest Olympus infrared laser-based z-drift compensation system (ZDC), which interferes with neither fluorescence imaging nor cell viability, is available for scanR to maximize autofocus accuracy, reliability, and speed. The ZDC perfectly complements the distinct autofocus options of the software to ensure the best focus accuracy and reliability.
Olympus Incubation System
The Olympus cellVivo provides stringent control of temperature, humidity, and CO2 levels for sample integrity and focus position when working with living cells.
The scanR team of application specialists will be happy to customize your system to suit your needs and applications precisely.
|ScanR screening system||
Microscope-based screening system platform for life science applications
Flexibility: system configuration can be adapted precisely to suit the application
Performance and endurance: a highly integrated system concept and real-time synchronization unites the advantages of an open
platform with the demands of screening applications for throughput and reliability
Olympus inverted high-end microscope IX83, one or two decks
Motorized stage, Märzhäuser SCAN IM 120 x 80 for IX83
Short-arc burners, 150 W, xenon or mercury/ xenon
Highly stabilized light output, intensity fluctuations < 0.1 %, feedback-loop-stabilized burner current
Fast filter wheel with 8 filter positions for standard 25 mm optical filters that are easy to exchange without tools
Attenuation, 14 levels from 1 % to 100 %, achromatic, high level of repeatability
Filter/attenuation switch <58 ms (adjacent positions)
Attenuation switch, min. 58 ms (adjacent positions)
Shutter, turn on/ off <1 ms
Lumencor Spectra X
6 independent LED channels, turn on/ off ~ 10 μs
Application-optimized bandpass filters
Lifetime > 20,000 hours with more than 70 % initial intensity
Transmitted-light illumination for visual inspection only (no transmitted-light screening)
Transmitted-light illumination for screening and visual inspection including fast shutter (transmitted-light screening supported)
Optional DIC (differential interference contrast) or phase contrast
Hardware control and
Real-time controller with additional CPU, independent of the OS of the imaging PC
Temporal resolution: 1 ms
Timing precision: < 0.01 ms
Hardware-synchronized multitask acquisition (illumination control, exciter filter, shutters, etc.)
Precise camera control via external trigger
Hamamatsu ORCA-FLASH 4.0 V2, high-sensitivity cooled sCMOS camera with large 18.8 mm sensor chip
Hamamatsu ORCA-FLASH 4.0 LT, economic sCMOS camera with large 18.8 mm sensor chip
Hamamatsu ORCA R2, high-sensitivity cooled CCD camera, recommended for long exposure times
Hamamatsu C8484, high-sensitivity CCD camera
Objectives for “thin” (0.1 - 0.2 mm) substrates, cover slips, and glass bottom plates (2x, 4x, 10x, 20x, 40x, 60x,100x)
Objectives for “thick” (~ 1 mm) substrates, plastic bottom plates, and slides (2x, 4x, 10x, 20x, 40x, 60x)
Phase contrast objectives for “thin” (0.1 - 0.2 mm) substrates, cover slips, and glass bottom plates (10x, 20x, 40x)
Phase contrast objectives for “thick” (~ 1 mm) substrates, cover slips, and glass bottom plates (10x, 20x, 40x)
Single-band filter sets (specifications as requested)
Multiband filter sets (specifications as requested)
2 independent software modules: ScanR Acquisition software and ScanR Analysis software
Shading correction workflow to compensate for shading and optimize spatial intensity homogeneity during and post-acquisition
The software modules can be installed on the same or on different workstations (Windows 7 32/64 bit)
Work-flow-oriented configuration and user interface
Variable, powerful software autofocus procedures that can be combined with an optional IR laser hardware autofocus function, 2-step coarse and fine autofocus, object-based autofocus, or image-based autofocus
Versatile plate manager with predefined formats (slides, multiwell plates) and editing interface to create and edit customized formats (spotted arrays)
Time-lapse screening, Z-stack screening, multicolor screening (unlimited number of acquisition channels)
User interaction: Pause, resume, reconfigure software autofocus
Support for integration into automated sample-preparation lines, e.g. scriptable interfaces for liquid handling
Automated image and data analysis for standard assays and assay development
Online and offline multicore analysis
Image processing, image analysis, particle detection, and parameter extraction and calculation
Cytometric data exploration, analysis, gating, and classification
Powerful and flexible gating concept including automated population detection
Direct link between data points, objects, and images
Assays-based work flow and advanced scientific assay development functionality
|Computer||Imaging computer (latest generation PC) , Windows 7 64 bit|
Time-lapse kinetic analysis module – a unique cytometric approach for better analyzing and understanding live cell dynamics
IR laser hardware autofocus function based on the ZDC of the latest Olympus IX83 microscope series
cellVivo incubation system
Encoded magnification changer IX3-CAS
Fast emission filter wheel (FFWO) for high-speed imaging in “Sedat” configuration
Customization: hardware, software, assays
Additional scanR Analysis workstation
|2 systems in one setup||Can be combined with cellSens live cell imaging software for full versatility of a high-end imaging system|