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Applied Optics

Applied Optics


  • Vol. 38, Iss. 4 — Feb. 1, 1999
  • pp: 714–719

Polarization effects in flow cytometric DNA sizing

Alexandra Agronskaia, Juleon M. Schins, Bart G. de Grooth, and Jan Greve  »View Author Affiliations

Applied Optics, Vol. 38, Issue 4, pp. 714-719 (1999)

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We have investigated the influence of the polarization direction of excitation light on DNA sizing results obtained by a flow cytometric technique. We found strong fluorescence anisotropy of the fluorescent signal from λ DNA stained with the bis-intercalating dye TOTO-1. Small fragments of DNA are less sensitive to polarization than larger pieces. This effect is more pronounced at faster flow speeds. These observations show a noticeable orientation of the DNA molecules introduced by the shear forces in the flow system. The data are consistent with an angle between the transition moment of fluorescence of TOTO-1, and the long axis of DNA is approximately 62°.

© 1999 Optical Society of America

OCIS Codes
(160.1190) Materials : Anisotropic optical materials
(260.2510) Physical optics : Fluorescence
(260.5430) Physical optics : Polarization
(280.2490) Remote sensing and sensors : Flow diagnostics

Original Manuscript: April 21, 1998
Revised Manuscript: July 20, 1998
Published: February 1, 1999

Alexandra Agronskaia, Juleon M. Schins, Bart G. de Grooth, and Jan Greve, "Polarization effects in flow cytometric DNA sizing," Appl. Opt. 38, 714-719 (1999)

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