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Effect of Fixative on Chromatin Structure and DNA Detection
Author(s) -
Falconi Mirella,
Teti Gabriella,
Zago Michela,
Pelotti Susi,
Gobbi Pietro,
Breschi Lorenzo,
Mazzotti Giovanni
Publication year - 2007
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.20440
Subject(s) - fixative , chromatin , interphase , paraformaldehyde , dna , in situ , in situ hybridization , glutaraldehyde , biology , microbiology and biotechnology , dna extraction , ultrastructure , chemistry , biophysics , biochemistry , chromatography , gene , cytoplasm , polymerase chain reaction , anatomy , gene expression , organic chemistry
In this study, we analyzed the chromatin ultrastructure in interphase cells after different chemical fixations. In light of the fact that there is little information regarding the fixation of biological samples in combination with molecular biology methods (such as DNA extraction and in situ hybridization methods) we analyzed the ultrastructure of chromatin in interphase cells fixed with different fixatives and tested under the same conditions for both DNA extraction and in situ hybridization. The results showed that, among the different combinations and concentrations we analyzed, the solution of 4% paraformaldehyde/0.1% glutaraldehyde was the best compromise in order to achieve a well‐preserved morphology, successful DNA extraction, and specific signaling of in situ hybridization, suggesting a low interference of this fixative with the chromatin organization. Microsc. Res. Tech., 2007. © 2007 Wiley‐Liss, Inc.