ISSN:
1573-6849
Keywords:
5-bromo-2′-deoxyuridine incorporation
;
chromosome structure
;
immunostaining
;
scanning ion microscopy
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract We investigated the possibilities of using scanning ion analytical microscopy (SIAM) to detect bromine in human metaphase chromosomes. The experiments were performed after incorporation of the thymidine analogue, 5-bromo-2′-deoxyuridine (BrdU), into the DNA or byin situ hybridization of a BrdU-labelled probe for the subcentromeric repeated DNA sequences. The possibilities offered by this microanalytical method were compared with immunofluorescent staining techniques. Well-defined maps of bands containing bromine were obtained with metaphase chromosomes that had incorporated BrdU during the late S-phase. Their patterns were similar to the labelling obtained by immunofluorescence. In addition, SIAM reveals the presence of bromine within constitutive heterochromatic regions in which BrdU is poorly detected by immunofluorescence. The comparison of the12C14N,31P and81Br maps of controls and fluorescence plus Giemsa (FPG) metaphase chromosomes, shows the loss of bromine from DNA during this treatment. SIAM emerges as a new powerful microanalytical technology for investigating chromosome structure further.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF02261720
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