THE MULTICOLOR FLUORESCENCE IN SITU HYBRIDIZATION (mFISH) HOMEPAGE
Liehr T*
*Corresponding Author: Dr. Thomas Liehr, Institut für Humangenetik und Anthropologie, Kollegiengasse 10, D-07743 Jena, Germany; Tel.: +49-3641-935533; Fax: +49-3641-935582;E-mail: i8lith@mti.uni-jena.de
page: 27

INTRODUCTION

The discipline of cytogenetics went through many stages of development, each providing more possibility for characterization of normal and structurally abnormal chromosomes [1]. Characterization of such chromosomes in humans is of high clinical importance and is the requisite condition for further molecular investigation aimed at identi- fication of disease-related genes [2,3] or positional cloning strategies [4]. Also, cytogenetic studies are significant for understanding the evolution of different animal species including Hominoids [5]. In the last 25-30 years, studies were accomplished by molecular cytogenetics, i.e., fluorescence in situ hybridization (FISH). In 1969, Gall and Pardue [6], introduced in situ hybridization, which, for the first time, allowed nucleic acid sequences to be examined inside cells or on chromosomes. However, because radioactivity has well-known shortcomings [7], the radioactive probe for labeling was replaced by alternative labeling modalities or detection systems, e.g., biotin detected by binding to avidin coupled to a fluorochrome [8]. Pinkel and coworkers [9] described the FISH technique that could be applied to human chromosomes and very soon thereafter the first twocolor FISH study was published [10]. We have defined multicolor FISH (mFISH) as the simultaneous use of at least three different ligands or fluo rochromes for the specific labeling of DNA (excluding the counter stain) [11]. According to this definition, the first successful mFISH experiments were those of Nederlof and coworkers [12] who visualized three differently-labeled nucleic acid sequences, simultaneously in blue [amino methyl coumarin acetic acid (AMCA)], in red [tetramethylrhodamine isothiocyanate (TRITC)] and in green [fluo rescein isothiocyanate (FITC)].Multicolor FISH assays have become indispensable for precise descriptions of chromosomal rearrangements, and studies including radiobiology, mutagenesis, inter-phase-architecture and chromosomal evolution. As the literature on this topic is huge, we created a regularly updated internet site in which all studies that apply mFISH are collected.




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