New technique for high resolution DNA sizing in epi-illumination


Schins, J.M. and Agronskaia, A. and Grooth, B.G. de and Greve, J. (1998) New technique for high resolution DNA sizing in epi-illumination. Cytometry Part A, 32 (2). pp. 132-136. ISSN 0196-4763

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Abstract:We present a high-resolution DNA-sizing technique based on the principles of flow cytometry, using a high numerical aperture objective and epi-illumination. The new technique, designed for small fluorescing samples/particles (sub-micron diameter) suspended in a weakly fluorescent medium, makes use of an additional focus for high-precision particle localisation. This way, only those particles are considered that flow exactly through a well-defined volume. Results are presented for fluorescent beads, as well as for YOYO-stained plasmids containing 5,500 basepairs. The latter were measured with 6.2% resolution, setting a new limit to flow-based sizing of DNA.
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Copyright:© 1998 Wiley-Liss
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