Near-Field Fluorescence Imaging of Genetic Material: Toward the Molecular Limit

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Hulst, N.F. van and Garcia-Parajo, M.F. and Moers, M.H.P. and Veerman, J.A. and Ruiter, A.G.T. (1997) Near-Field Fluorescence Imaging of Genetic Material: Toward the Molecular Limit. Journal of Structural Biology, 119 (2). pp. 222-231. ISSN 1047-8477

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Abstract:Chromosomes, DNA, and single fluorescent molecules are studied using an aperture-type near-held scanning optical microscope with tuning fork shear force feedback. Fluorescence in situ hybridization labels o­n repetitive and single copy probes o­n human metaphase chromosomes are imaged with a width of 80 nm, allowing their localization with nanometer accuracy, in direct correlation with the simultaneously obtained topography. Single fluorophores, both in polymer and covalently attached to amino- silanized glass, are imaged using two-channel fluorescence polarization detection. The molecules are selectively excited according to their dipole orientation. The orientation of the dipole moment of all molecules in o­ne image could be directly determined. Rotational dynamics o­n a 10-ms to 100-s timescale is observed. Finally, shear force imaging of double-stranded DNA with a vertical sensitivity of 0.2 nm is presented. A DNA height of 1.4 nm is measured, which indicates the nondisturbing character of the shear force mechanism.
Item Type:Article
Copyright:© 1997 Elsevier Science
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Link to this item:http://purl.utwente.nl/publications/23813
Official URL:http://dx.doi.org/10.1006/jsbi.1997.3888
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Metis ID: 128612