Direct visualization of individual DNA molecules by fluorescence microscopy: characterization of the factors affecting signal/background and optimization of imaging conditions using YOYO.

Gurrieri S, Wells KS, Johnson ID, Bustamante C
Anal Biochem. 1997 249 (1): 44-53

PMID: 9193707 · DOI:10.1006/abio.1997.2102

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.

MeSH Terms (8)

Acridine Orange Benzoxazoles DNA, Viral Ethidium Fluorescent Dyes Image Processing, Computer-Assisted Microscopy, Fluorescence Quinolinium Compounds

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