Metafer MetaCyte Multiparametric Cell Analysis

General

Brightfield MetaCyte fills the gap between fast, yet sometimes inflexible flow cytometers and microscope based imaging systems. MetaCyte combines MetaSystems' proven scanning platform Metafer with the latest multi color analysis and feature extraction algorithms. MetaCyte analyses cell features in single cell preparations and tissue sections, under fluorescent light conditions or in the brightfield mode. Sophisticated image processing algorithms cope with poor slide conditions or weak signals. Flexible cell selection based on the counter stain information is used to identify cells or tissue section regions that are suitable for analysis. More than 120 features Fluorescence per color channel, including intensity, morphology, and texture information, can automatically be extracted in real time. During the scan an image gallery of the analyzed objects is being built up. Histogram or scatter plot diagrams of selected features are displayed and updated on the fly. Interactive gating extracts exactly the subpopulation of cells that match the wanted criteria.

After the scan each individual cell can be checked on-screen or revisited under the microscope. Selection or rejection of images from the gallery and correction of automatic results is possible. All data obtained automatically during the scan is stored with the gallery images, can be printed into customizable reports, or may be exported into external spreadsheet or statistics software.

Metafer MetaCyte Options and Links
Due to its flexibility MetaCyte can be used for a variety of applications.	more...
MetaCyte automatically generates extended focus images.	more...
The AutoCapt module is an integrated part of MetaCyte.	more...
The Isis FISH imaging system can be seamlessly integrated.	more...
The CometScan module upgrades MetaCyte to an automatic Comet assay system.	more...
The slide feeder enhances the capacity of Metafer MSearch to 80 slides.	more...

Since MetaCyte is based on a flexible classifier concept, it can be adapted to various specimen conditions. MetaCyte's unique training feature allows the automatic optimization of classifiers. Based on a learning data set of interactively pre-classified images the software will find the criteria that give the best discrimination of the cells of interest and the best match of automatic and interactive classification.

As can be expected from a member of the Metafer systems familiy, MetaCyte provides ultra-high scanning speed. Depending on the application, MetaCyte scans and evaluates up to 500 cells per minute - absolutely unattended.

Applications

Due to its high flexibility and the unique training features MetaCyte is open for a variety of different applications (see examples below). In case you want to know if MetaCyte can help you with your own research or diagnosis project, please feel free to contact MetaSystems to ask for further details. Please refer also to our application section of this webspace.

Interphase FISH Signal Analysis (Spot Counting)

Automatic counting of FISH signals in a large number of interphase nuclei is a demanding task, since the signals are small and 3D-analysis is required due to the thickness of the nuclei. However, this application is widely used in cytogenetics and oncology, and automization bears many advantages. Besides the obvious benefit of automatic and unattended analysis, the use of Metafer MetaCyte generates a new quality of reproducibility and documentation. Usually FISH slides can not be kept for a long time period, and therefore there is no possibility to reproduce manual scoring results once the slides are no longer available. MetaCyte automatically creates a data set, that not only contains the evaluation of signals, but also an image of each single cell or region. Doubtful cases can be revised any time, and if necessary, the data may be corrected manually.

Spot Counting / Detection of Gene Fusions and Split Signals

With its ability to analyse 3D distances even between signals from different color channels MetaCyte is the perfect tool for automatic analysis of signal fusion or split signal assays. These assays are widely used in haematology (i.e., for the analysis of CML or CLL specimen) to detect characteristic chromosomal rearrangements, and many probe kits for various applications are commercially available. With its classifier concept MetaCyte can automatically score any of these specimen. After an extended focus image is generated, the signal features can be detected in combination with their spatial distribution, and fused signals can be counted. With the help of derived features (i.e., box classifiers) the cells may automatically be assigned to classes with objects showing the same phenotype. With this option it is very easy to determine how many cells provide certain staining patterns.

Automated detection of fusion signals. Left image:gallery image of a cell showing a fusion between on red and one green signal. Right image: cell from the same slide without fusion. Green numbers stand for the detected spot count in the green color channel, red numbers for the spot count in the red channel. The white number gives the minimal distance between a red and a green signal and is zero, if a fusion is present.

Cancer Diagnostics: Detection of Gene Amplification in Tissue Sections

The evaluation of gene amplification in oncogenes has become an important step of cancer diagnosis and therapy monitoring. MetaSystems provides with MetaCyte the tool for a reliable automatic detection of gene amplifications and deletions in paraffine embedded tissue sections and cytospin preparations, either in the brightfield mode or with fluorescence. Various cell sampling algorithms, including the new and revolutionary Tile Sampling method, provide precise detection of target cells. The evaluation of intensity values and areas from gene specific FISH signals, or from immunohistochemically stained cell compartments (including CISH staining patterns), leads to highly specific results providing the amplification status of the gene. The comprehensive data presentation allows easy selection of cell population subsets which show similar amplification rates. With the option to scan position lists, the researcher can interactively determine regions of interest in a tissue section (i.e., regions providing tumour material), and let Metacyte selectively scan only the pre-defined regions.

Toxicology: Micronucleus Assays / Comet Assay / CometFISH

Applications in genetic toxicology aim to precisely quantify DNA damage, and therefore require large sample sizes in order to achive statistically relevant data. The Metafer slide scanning platform with the module MetaCyte allows for automation of several toxicology assays on one system.

MetaCyte offers fast routines for automating both the rodent erythrocyte (fully OECD compliant) and the cytokinesis-block micronucleus assay. A barcode reading option, trainable detection rules, and the 80-slides SlideFeeder (available for all assays) increase throughput and quality at the same time. MetaCyte is fully GLP compliant, and is manufactured in Germany under rigid quality management.
MetaCyte's unique concept of user-trainable classifiers means it is adaptable to a virtually unlimited number of applications, including automated analysis of fluorescent signals in cells (e.g. the assessment of protein foci or FISH spot signal assays), the detection of rare cells at highest speed, or the analysis of tissue sections. Please contact us to find out if your application can also be automated with MetaCyte.

Forensics: Sperm Detection

Detection of sperm in crime scene samples is usually one important step in the process of confirming sexual assault in cases of rape. Automated sperm detection with Metafer MetaCyte offers a fast, yet highly reliable alternative to the tedious visual sample inspection, thus improving efficiency in solving crime cases.
MetaCyte identifies target cells by morphology and color, and displays them in a comprehensive color image gallery. A mouse click re-locates positions under the microscope for visual inspection. Up to 80 slides can be scored concurrently, and results of each sample are automatically stored together with the gallery images.

High Content Screening: Microwell Based Cell Screening

As cell assays in general and fluorescent cell assays in particular are becoming quite common in a number of applications, there is an increasing need for fast and efficient screening systems. High Content Screening (HCS) is a functional screening method based on high resolution fluorescence imaging of multiple targets in the context of intact cells. With the MetaCyte HCS option MetaSystems offers an automatic cell scanning system that meets all requirements for HCS: complete automation (walk-away operation), very high throughput, high flexibility, user-trainability for custom designable cell assays, and affordability.

Image gallery of a live cell screening in a 96 well plate (NRK cells, transfected with YFP fused cDNA, counterstained with Hoechst 33342) after sorting by total fluorescence in YFP channel; upper part of gallery shows cells with brightest fluorescence, lower part of gallery shows cells without YFP expression.

With its HCS abilities MetaCyte becomes a powerful tool for applications in molecular biology, cell biology, development, differentiation and molecular medicine. Connected to an inverted microscope and equipped with a special stage, MetaCyte can be used as a fully automatic screening system for HCS in microwell plates. Scanning a 96 microwell plate (2 color fluorescence, 4 fields of view per well at 20x optical magnification, which corresponds to typically 50 analyzed cells per well, 20 feature measurements per cell) just takes around 15 minutes.

Extended Focus Images

The cells or nuclei on a typical slide used for automated signal or morphology analysis usually is not a two-dimensional object. In many cases important information is distributed over various focus levels, and the snapshot of one selected focus plane would lock out critical features. Therefore MetaCyte is able to acquire a principally unlimited number of images per signal channel, that are captured in different focal planes. Each image may be enhanced individually, and the software automatically generates an extended focus image. This is done using an algorithm, that not just adds various focus levels to one image, but selectively transfers regions, that are in focus, into the final image. This method allows to determine 3D-distances between signals, and i.e. exclude coincidentally co-localized spots from real spot fusions.

The automatic generation of extended focus images
(click the image to see details).

Extended Focus Image Function (click to enlarge)

AutoCapt: Automatic Acquisition of High Magnification Images

The automated image acquisition software AutoCapt captures high magnification (i.e., 63x or 100x) images from cells detected by MetaCyte. It uses the position list which was generated by MetaCyte to relocate the cells, automatically centres and focusses the images and stores them ínto the MetaCyte data file. Images may be captured in the transmitted light and in fluorescence mode, the latter with up to six color channels. Like all Metafer applications, AutoCapt uses classifiers to adjust the way, how the images are captured. Once the images are generated, they can be exported into various file formats like TIFF, JPEG or BMP. Hereby it is possible to export all images at the same time, or to select subsets or even single images to be exported.
For networked installations, with Isis FISH imaging workstations connected to the MetaCyte system, it is even possible to let AutoCapt export its images into the proprietary file formats of these softwares. Images generated in this way may be easily opened on any external Isis workstation for further analysis.

AutoCapt is integrated into Metafer MetaCyte.

Integration of FISH Imaging

The Isis FISH imaging system seamlessly integrate into the MetaCyte system, and can be used to acquire high magnification, high quality images of the detected cells or nuclei. Of course Isis, like MetaCyte, also is capable to capture various focus planes and to generate extended focus images. Since all MetaSystems products are based on a modular architecture, many different installations are conceivable:

As a basic solution Isis may be installed on the Metafer system amd launched directly clicking a button in MetaCyte. With this configuration Metafer transfers data like case name, slide designation and coordinates into the FISH imaging software. Isis may also be integrated as capture module, while the image preparation is done on external workstations.
If the MetaCyte system is too busy to allow the use of Isis on the same station, the MetaCyte data may be opened on external relocation system (Relosys). Relosys workstations are equipped with a single slide motorized stage and provide a similar screen like the MetaCyte system itself. Like in Metacyte, cells may be relocated and subsequently captured and analyzed in Isis.
Since MetaCyte is extended with the AutoCapt software, Isis images may also be generated automatically. External workstations with access to the Metafer system amy then load the images for further analysis.
If there is no network connection between the Metafer system and the Isis workstations, the coordinates may also be converted to the coordinates of each microscope. Relocation can then be done manually using the microscope rulers.

However the final network will look like: all MetaSystems installations will be tailored to the needs of the user, and all products work seamlessly together.

The CometScan module extends the MetaCyte capabilities to automated Comet assay analysis

With the optional module CometScan any MetaCyte system can be upgraded to an automatic scanning system for single cell gel electrophoresis slides (Comet assay). CometScan automatically identifies cells on the slide and evaluates relevant parameters like head and tail lengthes and tail moments. Since CometScan is an integrated part of MetaCyte, the advantages of both softwares may be combined to do automated Comet FISH analyses.

For more details please refer to our CometScan page.	more...