Effect of fixative on chromatin structure and DNA detection.
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.
Falconi
Mirella
M
Department of Anatomical Sciences, University of Bologna, Italy.
Teti
Gabriella
G
Zago
Michela
M
Pelotti
Susi
S
Gobbi
Pietro
P
Breschi
Lorenzo
L
Mazzotti
Giovanni
G
D002460:Cell Line; D002843:Chromatin / Q000648:ultrastructure*; D004247:DNA / Q000648:ultrastructure*; D005404:Fixatives; D005976:Glutaral; D006652:Histological Techniques; D006801:Humans; D017403:In Situ Hybridization / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D016707:Tissue Fixation / Q000379:methods*; D014407:Tumor Cells, Cultured
D002843:Chromatin; D005404:Fixatives; D004247:DNA; D005976:Glutaral