OSA's Digital Library

Applied Optics

Applied Optics

APPLICATIONS-CENTERED RESEARCH IN 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)
http://dx.doi.org/10.1364/AO.38.000714


View Full Text Article

Acrobat PDF (369 KB)





Browse Journals / Lookup Meetings

Browse by Journal and Year


   


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools

Share
Citations

Abstract

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

Citation
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)
http://www.opticsinfobase.org/ao/abstract.cfm?URI=ao-38-4-714


Sort:  Author  |  Year  |  Journal  |  Reset

References

  1. P. M. Goodwin, M. E. Johnson, J. C. Martin, P. W. Ambrose, B. L. Marrone, J. H. Jett, and R. A. Keller, “Rapid sizing of individual fluorescently stained DNA fragments by flow cytometry,” Nucl. Acids Res. 21, 803–806 (1993).
  2. A. Castro, F. R. Faifield, and E. B. Shera, “Fluorescence detection and size measurement of single DNA molecules,” Anal. Chem. 65, 849–852 (1993).
  3. J. T. Petty, M. E. Johnson, P. M. Goodwin, J. H. Jett, and R. A. Keller, “Characterization of DNA size determination of small fragments by flow cytometry,” Anal. Chem. 67, 1756–1761 (1995).
  4. A. Castro and E. B. Shera, “Single-molecule detection: applications to ultrasensitive biochemical analysis,” Appl. Opt. 34, 3218–3222 (1995).
  5. Z. Huang, J. T. Petty, B. O’Quinn, J. L. Longmire, N. C. Brown, J. H. Jett, and R. A. Keller, “Large DNA fragment sizing by flow cytometry: application to the characterization of P1 artificial chromosome (PAC) clones,” Nucl. Acids Res. 24, 4202–4209 (1996).
  6. H. S. Rye, S. Yue, D. E. Wemmer, M. A. Quesada, R. P. Haugland, R. A. Mathies, and A. N. Glazer, “Stable fluorescent complexes of double-stranded DNA with bis-intercalating asymmetric cyanine dyes: properties and applications,” Nucl. Acids Res. 20, 2803–2812 (1992).
  7. A. Larsson, C. Carlsson, M. Jonsson, and B. Albinsson, “Characterization of the binding of the fluorescent dyes YO and YOYO to DNA by polarized light spectroscopy,” J. Am. Chem. Soc. 116, 8459–8465 (1994).
  8. C. Carlsson, A. Larsson, M. Jonsson, B. Albinsson, and B. Norden, “Optical and photophysical properties of the oxazole yellow DNA probes YO and YOYO,” J. Phys. Chem. 98, 10313–10321 (1994).
  9. V. Kachel, H. Fellner-Feldegg, and E. Menke, “Hydrodynamic properties of flow cytometry instruments,” in Flow Cytometry and Sorting, 2nd ed., M. R. Melamed, T. Lindmo, and M. L. Mendelsohn, eds. (Wiley, New York, 1990), pp. 27–44.
  10. A. Agronskaia, A. Florians, K. O. van der Werf, J. M. Schins, B. G. de Grooth, and J. Greve, “A photon counting device compatible with conventional flow cytometric data acquisition electronics,” Cytometry 32, 255–259 (1998).
  11. T. C. Bakker Schut, A. Florians, K. O. van der Werf, and B. G. de Grooth, “Flow cytometry signal processing and data acquisition with a personal computer using an RTI-800 multifunction A/D I/O board,” Rev. Sci. Instrum. 64, 3116–3120 (1993).
  12. B. Norden, M. Kubista, and T. Kurucsev, “Linear dichroism spectroscopy of nucleic acids,” Q. Rev. Biophys. 25, 51–170 (1992).

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.


« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited