Multicolor Imaging

Fluorescent Chromosome Analysis

The Ikaros Karyo C module for multicolor FISH (mFISH) and multicolor chromosome banding (mBAND).

Multicolor FISH assays are used for a precise assessment of complex chromosomal rearrangements. The assays are based on combinations of fluorochromes which are interpreted by the analysis software. The method of combinatorial labeling effectively multiplies the number of usable labels which would otherwise be limited to the 7 or 8 distinguishable individual fluorochromes.

Multicolor FISH

Multicolor FISH (mFISH) is a method to facilitate analysis of each single chromosome or chromosome part of a metaphase. Thus, marker chromosomes, complex chromosomal rearrangements, and all numerical aberrations can be visualized simultaneously in a single hybridization experiment. By analyzing the color ratios of all pixels, Ikaros Karyo C unambiguously assigns any region of the image to the respective chromosome class. The filter based mFISH approach facilitates a direct inspection of the original hybridization signals and, thus, overcomes the need for additional control experiments with conventional chromosomal paints.

Versatile tools for precise chromosome separation and for rapid chromosome classification help speeding up the workflow. Chromosome classification is either done based on DAPI banding or by using the fluorochrome combinations. mFISH chromosomes can be displayed in three different pseudo color display modes in order to facilitate the analysis: maximum saturation (solid color), DAPI image overlaid with the pseudo color, and standard pseudo color image.

Analysis of mFISH images in Ikaros Karyo C is easy even if complex aberrations are involved. The Pixel Info / Extended Pixel Info tools display the chromosome class assigned to the selected pixel. The reliability index of the assignment can also be displayed. The False Color Binary Display virtually transforms the mFISH image into a series of hybridizations with 3 individual chromosome paints. This tool was introduced to facilitate the analysis of mFISH images with numerous chromosome rearrangements that cannot be identified "at a glance". Ambiguous color signatures can optionally be excluded from the classification. This way, regions with insufficient hybridization are easily identified and suppressed. The Single Color Gallery is a separate window, in which all five color channels are displayed for a selected chromosome. Additionally, the false color image, an inverted DAPI image, and the fluorochrome profiles are shown. With the help of the Single Color Gallery, the identification of rearrangements is extremely easy and straightforward.

Need assistance?

Multicolor Banding

Multicolor chromosome banding (mBAND), a proprietary technique of MetaSystems Probes, principally uses the technique of multicolor FISH (mFISH) and applies it to the detection of intra-chromosomal rearrangements. In detail, the probes (XCyte mBand probes) labeled with different proportions of two or more fluorochromes are hybridized to a single chromosome pair. Each probe consists of overlapping partial chromosome paints. The changing fluorescence intensity ratios along the chromosomes are used to assign different pseudo-colors to specific chromosome regions in the Ikaros Karyo C software. The result is a reproducible high-resolution false-color banding pattern allowing for identification and localization of intra-chromosomal aberrations.

The resolution of the mBAND pattern can be modulated in order to get the most suitable image for analysis. A resolution equivalent to 550-band level for G-bands can be easily achieved. mBAND patterns are independent of chromatin condensation. Though mBAND bands resemble an artificial banding pattern, they can be directly correlated with ISCN bands using the single color gallery Ikaros Karyo C. Hence, precise localization of breakpoints is possible.

Need assistance?


Legal Note

MetaSystems software and system products are classified as in vitro diagnostic medical devices (IVD) in the European Union in accordance with In Vitro Diagnostics Regulation (EU) 2017/746 or In Vitro Diagnostic Medical Device Directive 98/79/EC, respectively, and carry the CE label unless otherwise indicated. Use all MetaSystems IVD products only within the scope of their intended purpose.

MetaSystems products are used in many countries worldwide. Depending on the regulations of the respective country or region, some products may not be used for clinical diagnostics.

Some hardware components supplied by other manufacturers are not included in MetaSystems IVD products and are therefore not IVD medical devices.

MetaSystems software provides, among other functions, features to assist users with image processing. These include, but are not limited to, the use of machine and deep learning algorithms for pattern recognition. The output generated in this process should be regarded as preliminary suggestions and, in any case, mandatorily requires review and assessment by trained experts.

MetaSystems offers Customization Packages for application workflows that have been successfully implemented for customer labs using standard Metafer platform functionality. It is expected that they can be implemented for other customer labs using similar workflows and slide preparation procedures. If a Customization Package is purchased, MetaSystems product specialists will - based on their experience from other similar application cases - support the customer lab in adapting the Metafer software configuration to their needs. The performance of the solution will depend on the quality of the customer slides and the expertise of the users, MetaSystems cannot specify or guarantee any performance parameters. The validation of the solution for clinical use is the sole responsibility of the customer lab.

Please contact us for further information.