Automatic detection of stable and unstable chromosome aberrations visualized by three‐color imaging after fluorescence in situ hybridization with a painting and a pancentromeric DNA probe

Abstract

Two-color fluorescence in situ hybridization (FISH) in combination with digital image analysis was used to develop an automatic system for the detection and classification of chromosome aberrations. Algorithms were developed for the automatic thresholding of the three digitized images: an FITC image representing specific painted chromosomes, a TRITC image representing the centromeres of all chromosomes, and a DAPI image representing all the counterstained chromosomes. A further algorithm was developed for the automatic classification of the different types of chromosome aberrations, such as translocations, dicentrics, and fragments. For this study, a dataset of 252 metaphases were digitized and analyzed automatically as well as manually. Of these metaphases, 81.3% could be correctly classified by the algorithm. The error rate was reduced to 9.3% by automatically excluding the detected clusters and artifacts. The average analysis time per metaphase was 34.5 s without any user intervention.

Use of an Automated Image Processing Program to Quantify Recombinant Adenovirus Particles

“…The approach has become a valuable analytical tool within the field of microscopy as the number of applications has increased and instruments have become more sophisticated with improved precision of measurement. Semiautomatic and completely automated image processing methods have been shown to be useful in biological science and medicine~Gatlin et al, 1993;Nelson et al, 2000;Swedlow et al, 2003!. For example, the automation of chromosome aberration scoring has reduced the burden of routine chromosome analysis for clinical diagnostics, thus allowing for unambiguous data acquisition and increased sample throughput~Coco- Martin et al, 1998!. For radiological studies, automated processing has been particularly useful in assisting in the viewing and interpretation of digital radiographs, such as magnetic resonance images of the brain and radiograph images of the chest and extremities~Huh Koenker & Grover, 2002!. Digital imaging and processing has also created a driving interest in the use of microscopy for large-scale screening using informatics for the storage and analysis of microscope image data. For example, stored data sets of small molecules such as gene sequences along with related digital image recordings can be accessed in open databases, thereby bridging the gap between sequence and function~Mayer et al The implementation of digital imaging into the field of electron microscopy~EM!…”

Toward an automated system for the analysis of cytogenetic abnormalities using fluorescence in situ hybridization technique

“…Most of these approaches are based on threshold grey level histogram techniques, either with no further analysis, when the grey level contrast is sufficiently high [2], or with an analysis of the area distribution of the stained chromosomes [3]. Some approaches use centromere staining in addition to chromosome hybridization [4]. Encouraging results have been obtained by using a more sophisticated analysis which however requires an expensive hybridization technique such as multiplex-FISH (M-FISH) [5].…”