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Direct Visualization of Individual DNA Molecules by Fluorescence Microscopy: Characterization of the Factors Affecting Signal/Background and Optimization of Imaging Conditions Using YOYO

1997; Elsevier BV; Volume: 249; Issue: 1 Linguagem: Inglês

10.1006/abio.1997.2102

1096-0309

Sergio Gurrieri, K. Sam Wells, Iain Johnson, Carlos Bustamante,

Monoclonal and Polyclonal Antibodies Research

A new series of high-affinity cyanine dyes was tested for the visualization of the dynamics of single DNA molecules through a fluorescence microscope. In particular, YOYO-1 (1,1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)- bis-4-[3-methyl-2,3-dihydro-(benzo-1,3-oxazole)- 2-methylidene]-quinolinium tetraiodide) forms a very stable, highly fluorescent complex with double-stranded DNA and dramatically improves the quality of the images. We have characterized the factors affecting the signal/background in the imaging of single DNA molecules by fluorescence microscopy and compared the results obtained using YOYO-1 with those obtained using standard fluorescent dyes like ethidium bromide or acridine orange.