caa a22 4500 463209796 CHVBK 20180405153137.0 cr unu---uuuuu 170326e20070401xx s 000 0 eng 10.1007/s00412-006-0084-2 doi (NATIONALLICENCE)springer-10.1007/s00412-006-0084-2 Preservation of large-scale chromatin structure in FISH experiments [Elektronische Daten] [Claudia Hepperger, Simone Otten, Johann von Hase, Steffen Dietzel] The nuclear organization of specific endogenous chromatin regions can be investigated only by fluorescence in situ hybridization (FISH). One of the two fixation procedures is typically applied: (1) buffered formaldehyde or (2) hypotonic shock with methanol acetic acid fixation followed by dropping of nuclei on glass slides and air drying. In this study, we compared the effects of these two procedures and some variations on nuclear morphology and on FISH signals. We analyzed mouse erythroleukemia and mouse embryonic stem cells because their clusters of subcentromeric heterochromatin provide an easy means to assess preservation of chromatin. Qualitative and quantitative analyses revealed that formaldehyde fixation provided good preservation of large-scale chromatin structures, while classical methanol acetic acid fixation after hypotonic treatment severely impaired nuclear shape and led to disruption of chromosome territories, heterochromatin structures, and large transgene arrays. Our data show that such preparations do not faithfully reflect in vivo nuclear architecture. The Author(s), 2006 Hepperger Claudia Department Biologie II, Ludwig-Maximilians-Universität München, Großhaderner Str. 2, 82152, Planegg-Martinsried, Germany aut Otten Simone Department Biologie II, Ludwig-Maximilians-Universität München, Großhaderner Str. 2, 82152, Planegg-Martinsried, Germany aut von Hase Johann Kirchhoff Institut für Physik, Universität Heidelberg, Heidelberg, Germany aut Dietzel Steffen Department Biologie II, Ludwig-Maximilians-Universität München, Großhaderner Str. 2, 82152, Planegg-Martinsried, Germany aut Chromosoma Springer-Verlag 116/2(2007-04-01), 117-133 0009-5915 116:2<117 2007 116 412 https://doi.org/10.1007/s00412-006-0084-2 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00412-006-0084-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hepperger Claudia Department Biologie II, Ludwig-Maximilians-Universität München, Großhaderner Str. 2, 82152, Planegg-Martinsried, Germany aut NATIONALLICENCE 700 1- Otten Simone Department Biologie II, Ludwig-Maximilians-Universität München, Großhaderner Str. 2, 82152, Planegg-Martinsried, Germany aut NATIONALLICENCE 700 1- von Hase Johann Kirchhoff Institut für Physik, Universität Heidelberg, Heidelberg, Germany aut NATIONALLICENCE 700 1- Dietzel Steffen Department Biologie II, Ludwig-Maximilians-Universität München, Großhaderner Str. 2, 82152, Planegg-Martinsried, Germany aut NATIONALLICENCE 773 0- Chromosoma Springer-Verlag 116/2(2007-04-01), 117-133 0009-5915 116:2<117 2007 116 412 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer