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Scanning ion analytical microscopy for high-resolution detection of 5-bromo-2′-deoxyuridine incorporation in metaphase chromosomes (1996)

Bourgeois, C. A. ; Dennebouy, R. ; Gibaud, A. ; [et al.]

Springer

Chromosome research 4 (1996), S. 574-582

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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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Fluorescence decay of ethidium bromide in the presence of the Z-conformation of poly-(dG-dC) and of poly(dG-dC) modified by chlorodiethylenetriamine platinum(II) chloride (1986)

Genest, D. ; Malfoy, B.

New York : Wiley-Blackwell

Biopolymers 25 (1986), S. 507-518

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ISSN: 0006-3525

Keywords: Chemistry ; Polymer and Materials Science

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Chemistry and Pharmacology

Notes: A time-resolved fluorescence study of ethidium bromide (EB) in the presence of poly(dG-dC) and of poly(dG-dC) modified by chlorodiethylenetriamine platinum(II) chloride is presented under solvent conditions in which these polymers adopt the Z-conformation (high ionic strength). It is shown that these polynucleotides can intercalate a very small quantity of EB. The binding parameters have been determined. The fluorescence lifetime of EB is slightly higher when bound to the Z-conformation (≃25 ns) than when bound to the B-conformation (≃23.7 ns). The nature of the salt has been checked. In the presence of 2.5M NaClO4, no transition from the Z-conformation to another conformation is observed when EB is added. On the contrary, in the presence of 4.25M NaCl, EB induces a cooperative transition from the Z-conformation to a conformation characterized by a much higher affinity for EB intercalation. In the case of poly(dG-dC) this last conformation is identical to the one observed at low ionic strength (B-conformation), but in the case of the platinated polymer this conformation is slightly different, as judged by the smaller value of the fluorescence lifetime of the intercalated EB.

Additional Material: 4 Ill.