In Situ Hybridization to Chromosomes Stabilized in Gel Microdrops

Cytometry, 1995, vol. 21, pp. 111-119

Bao-Tram Nguyen, Kristina Lazzari, Joseph Abebe, Ivan Mac, James B. Lin, Albert Chang, Karen L. Wydner, Jeanne Bentley Lawrence, L. Scott Cram, Heinz-Utrich Weier, James C. Weaver and David W. Bradley

Conventional chromosome in situ hybridization procedures rely on fixation to glass slides followed by microscopic evaluation. This report describes the development of a microdrop in situ hybridization (MISH) method which facilitates hybridization to chromosomes in suspension. Chromosomes encapsulated in gel microdrops (GMDs) composed of an agarose matrix withstood stringent hybridization and denaturation conditions. Because of the increased stability, hybridization to encapsulated chromosomes was detected by flow cytometry as well as conventional microscopy. Thus, the MISH method offers a means for chromosome hybridization without slides and may enable identification and isolation of chromosomes using hybridization rather than nucleic acid binding dyes.

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