Specifications of the Thermo Scientific CellInsight Personal Image Cytometer

Transcription

1 The Thermo Scientific CellInsight Personal Image Cytometer is an integrated hardware/software and informatics solution for developing High Content Assays (HCA). It is a fully automated and integrated fluorescence-imaging platform and includes a complete suite of software and data management solutions for automated cell analysis, drug discovery, and toxicology. The system uses a custom built optical train developed and built by Thermo Fisher Scientific. Users of Thermo Scientific HCA platforms have published more than 400 peer-reviewed publications. The system uses automated image acquisition along with non-scripting pre-written applications/modules for real time analyses. Data generation, data visualization, data storage, data management, data manipulation and data archiving are seamlessly integrated as a complete platform. Specifications of the Thermo Scientific CellInsight Personal Image Cytometer The CellInsight Personal Image Cytometer platform approximates the size of a laser printer, does not require special electrical inputs, and can be installed by the customer. Optical Train: A custom inverted epifluorescence optical train with cooled CCD camera High Quality Image Acquisition Objectives: Olympus 10X Plan NeoFluar or a 20X Achroplan as standard components. Resolution: Example, 20x nm per pixel maximum resolution Stage, plate and slide compatibility: The CellInsight Personal Image Cytometer has an automated stage with plate insert that is suitable for scanning multi-well plates. All clear bottom plates with Standard SBS footprint can be used in the system. The system is capable of imaging glass as well as plastic flat clear bottom wells ranging anywhere between slides, 8 well chamber slides, 6 well plates, 12 well, 24 well, 48 well, 96 well, 384 well plates and 1536 wells as well as 35mm glass bottom petri dishes, labtek chambered cover glass, slides, or other custom plate and slide configurations. The system is capable of adjusting to any plate manufacturer type that fits SBS standards. There is a Calibration wizard that allows easy set up of plate dimensions to allow easy scans. Light Source: The system uses an LED light engine with four-color capability. With solid state fourcolor excitation capability, the LED Light Engine reduces exposure times, data variability, and consumable expenses as compared to traditional arc lamp technologies. Built specifically for imaging in the blue (Hoechst/DAPI), green (FITC/GFP), orange red (TRITC) and far red (Cy5) ranges, the LED light engine is perfect who want even more robust, reproducible, and fast results from their highcontent platform. The LED light engine has a life expectancy of greater than 50,000 hours. The LED source is on only when required (i.e. when running exposures during a scan). The LED Light Engine is completely compatible with all hardware modules and Thermo Scientific BioApplications using a combination of the four common wavelength areas. When compared to Arc Lamp technology, the LED light engine performs better in all common wavelengths with superior

2 performance especially in the red and far-red regions of the spectrum. This can significantly reduce the overall scan times in highly multiplexed assays. Emission filters: Four channel dichroic/emission filter pair allows multiplexing of up to 4 channels simultaneously. Multiplexed Channels: Using the CellInsight Personal Image Cytometer, up to six channels can be acquired sequentially per image. Background correction is available in every channel to allow accurate measurements. Analysis fully integrated across all channels in an image-set. Speed: The CellInsight Personal Image Cytometer takes about minutes per 96 well plate based on two-color and one field per well, with the ability to view cell and well feature data and export data and images as soon as the plate finishes scanning. You can also view and export the cell feature and well feature data for an imaged field in an interactive Excitation Wavelength Fluorophore Scan Plates Per Assays Per Time Day* Day# 386nm23/485nm20 Hoechst/Alexa488 3: , nm23/549nm15 Hoechst/Rhodamine 4: , nm23/650nm13 Hoechst/Alexa660 5: ,080 window prior to scanning the plate. All scans include real time data analyses, so that you get to see the data immediately after the scan is complete. Camera: The CellInsight Personal Image Cytometer, comes with a high-resolution, shutterless thermoelectrically cooled 12 bit (4096 greyscales) CCD camera with a chip size of 1024 x 1024 pixels. Images can be binned from 1 x 1 to 2 x 2 and 4 x 4 to save server space. The camera uses electronic shuttering. Normal resolution generates images that are around 500kB, which are compressed another 30% prior to spooling. Software The system has software with intelligent acquisition that allows you to define the number of cells of a particular phenotype (for example neuronal cells) within fields per well as required and also define sparse wells to perform real time analysis during image acquisition. Images are processed simultaneously with image acquisition, allowing machine intelligence to be built in for stopping criteria and real phenotypic analysis. This is useful to minimize time spent scanning sparse wells, and also to ensure a statistically significant number of cells are captured per well i.e. to ensure consistent Z. An additional benefit of real time analysis during acquisition is that the user can see the results of the scan immediately, without a need for subsequent, manual processing. No other major vendor software has such a configuration. Our software is also optimized to run on one PC unlike other HCS vendors who require distributed processing over multiple CPUs to increase throughput. The software allows setting exposure times in required dynamic ranges and also set reference wells for statistical analysis. The software also contains a built in event wizard that uses Boolean logic statements to address sub-population analysis. Recently, two new interfaces were launched for assay development and data analyses named Thermo Scientific idev and iview.

3 idev The new Thermo Scientific idev software is part of High Content 2.0, the next generation of hardware and software. idev software is a truly immersive, assay development experience providing a guided workflow with visual feedback on multiple biological states in parallel. Interactive slider bar controls allow users to modify parameters and see changes in real time, displayed at both the image processing and resulting data levels. idev simplifies creating and optimizing your High-Content assays and increases productivity. Features of idev include guided workflows for assay protocol setup, image acquisition, analysis and scanning. idev has a streamlined user interface that organizes parameters by function, channel, etc. A Digital microscope mode allows you to interact with your sample. Multiple groups of images (representing biological states to be addressed) can be collected and processed simultaneously. idev includes simplified manipulation of analysis parameters using radio buttons and slider bars for instant feedback on changes, real-time, interactive visualized results at both image processing and cell data level and statistics driven identification of the most relevant output features. Because of its ease of use, idev drastically reduces the learning curve for new and occasional users resulting in less training especially in multi-user environments. The intuitive interface organization results in a more logical approach when developing multiplexed assays. It allows easy set up and image acquisition in minutes. With idev, no specialist-computing infrastructure required, reducing the total cost of ownership compared to other systems that utilize grid, or multi-processor arrays. i-view Thermo Scientific iview offers an integrated workflow for data and image centric decisions. With iview you can Quickly QC your assay at the plate level to ensure proper labeling and plating, easily drill down from Plate View to Well, Field, and Cell levels to reveal meaningful changes in cell responses. i-view also lets the user Create and control kinetic movies from your live cell experiments and links back to plate scanning for easy preview or assay optimization tasks. Some of the screen shots for iview are shown below.

4 Multiple well display Cell level scatter plot with cell cut outs Phenotypic Gallery Multiple fields per well display User controlled brightness and contrast, Display and configure algorithm overlays, Zoom Tools, Cell level graphing with cell selection tools, Integrated online help and tutorials and Multiple field viewing capabilities to name a few. Both idev and iview will be available installed in the purchased instrument. The CellInsight Personal Image Cytometer comes with analysis software containing 16 Thermo Scientific BioApplication software modules. These Bioapplications are flexible validated modules that allow for real time analysis including cell population based on a particular phenotype and differentiating it from other phenotypes leading to more statistically relevant data. The algorithms automatically calculate a population of cells and report the data as a percent of control as it is not sufficient to only look at raw cell and well calculations. The BioApplications allow image processing which requires no coding of algorithms to process a batch of images and is validated to achieve robust results on multiple Biologies. The 16 BioApplications include: 1. Cell Cycle v4.0 BioApplication 2. Cell Health Profiling v3.0 BioApplication 3. Cell Motility v4.0 BioApplication 4. Cell Spreading v4.0 BioApplication 5. Colocalization v4.0 BioApplication 6. Compartmental Analysis v4.0 BioApplication 7. Cyto-Cell Membrane v4.0 BioApplication 8. Cyto-Nuc Translocation v4.0 BioApplication 9. GPCR Signaling v4.0 BioApplication 10. Molecular Translocation v4.0 BioApplication

5 11. Morphology Explorer v4.0 BioApplication 12. Multi-parameter Cytotoxicity v4.0 BioApplication 13. Neuronal Profiling v4.0 BioApplication 14. Spot Detector v4.0 BioApplication 15. Target Activation v4.0 BioApplication 16. Tube Formation v4.0 BioApplication The Thermo Scientific vhcs View data viewer allows visualization of data and images immediately after a plate gets scanned. Data generated can be visualized as presentation ready line graphs / bar charts as well as numbers that can be exported into Excel with a single click. vhcs View displays well, field as well as cell feature data that is exportable as excel files. The cells can be visually linked to data points in scatter plots allowing users to make intelligent decisions by excluding cell populations by spotting and excluding outliers thereby improving their Z scores and assay windows. Thermo Scientific vhcs View and vhcs Scan cross talk and allow images acquired to be reloaded into the vhcs Scan software permitting reanalysis of acquired images. Montage Tool: A montage tool allows users to montage scanned fields and reanalyze a larger field of view. This helps with analysis of biologies like large stem cell colony imaged with the instrument. Data Storage and Data Management The Thermo Scientific Cellomics Store Standard Edition (SE) allows automated addressing and loading of data from a HCS reader into a SQL Server or Oracle database. Store SE offers rapid, highly scalable, global access to all HCS data. Images and derived data are linked, allowing drill-down to the level of individual cells. The Standard Edition permits the connection of one licensed HCS instrument and includes one (1) server side license for the HCS Applications Server. Store SE supplies a comprehensive, fully linked database, allowing click-through from results to data to images and even to individual cells on images. Scan and View software are interlinked to allow easy rescanning of images. Some of the highlights include: Images and data are automatically transferred to an enterprise- or academic-class database. Without this, manual data storage would require human interaction to transport several CD ROMs per experiment. Centralized data storage solution allows data and image access from multiple locations. Images and data are accessible. This allows a scientist to access images that are linked to the derived data points. Images and data are linked. The Thermo Scientific data visualization software (vhcs : View) provides rich visualization of HCS data, allowing users to see spatial, temporal, morphological and phenotypic changes at the cell level. Users can also interactively, visually analyze multi-feature, cell-based data down to the image all the data is seamlessly linked together. For example if the user wants to see data for a particular cell, they need only to click on the cell in the image to see all the relevant data. Without the vhcs:view module, a scientist would have to manually link up plate, well and cell data with images in order to be able to see the complete data set.

6 More importantly If the acquired image format from any source, is a compatible one (i.e, C01, DIB, TIFs, BMPs), they can be seamlessly recognized thro the updated software thereby facilitating reanalysis using current more powerful bioapplication modules. Training Upon initial installation up to four people will be trained by an application specialist, who will also be responsible for subsequent support to anyone who uses the platform. The application support is unparalleled. Dedicated Thermo Scientific Application Scientists are distributed globally. Application Scientists do regular on-site visits to work with users on assay development, deliver seminars on HCA related biologies and also do remote WebEx sessions to ensure that they are users can maximize their HCA experience. In addition, we offer new-user and expert weeklong training courses in Pittsburgh, Pennsylvania, a bulletin board for user questions and telephone triage service. We also regularly organize user-group meetings to foster a community of users who can then learn from each other. In addition, Pittsburgh-based scientists are available for custom assay development assistance. Service and Technical Support and Response Time A dedicated service group services Thermo Scientific HCA instruments, with field and telephone agents. Almost all issues can be resolved on-site, and it is very rare for broken components to need shipping to Pittsburgh. Service ticketing and immediate telephone responses are handled out of Pittsburgh. There is 24-hour turn around time for service issues. A toll-free technical support hot line as well as is available for troubleshooting. Customers can talk live to a technical support person who will trouble shoot, provide remote assistance as well as triage customer queries according to the nature of the call. BN