Direct measurement of single-molecule diffusion and photodecomposition in free solution

Science

Volumn 275, Issue 5303, 1997, Pages 1106-1109

  Xu, Xiao Hong ^a   Yeung, Edward S ^a  

^a IOWA STATE UNIVERSITY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

RHODAMINE 6G; SINGLE STRANDED DNA;

ARTICLE; COVALENT BOND; DIFFUSION; FLUORESCENCE; MEASUREMENT; MOLECULAR DYNAMICS; PHOTODEGRADATION; PRIORITY JOURNAL; STOCHASTIC MODEL;

DIFFUSION; DNA, SINGLE-STRANDED; FLUORESCENCE; FLUORESCENT DYES; MICROSCOPY, VIDEO; PHOTOLYSIS; RHODAMINES; SOLUTIONS;

EID: 0031037896     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.275.5303.1106     Document Type: Article

References (29)

1. B. Rotman, Biochemistry 47, 1981 (1961); Q. Xue and E. S. Yeung, Nature 373, 681 (1995).
2. B. Rotman, Biochemistry 47, 1981 (1961); Q. Xue and E. S. Yeung, Nature 373, 681 (1995).
3. N. J. Dovichi, J. C. Martin, J. H. Jett, R. A. Keller, Science 219, 845 (1983).
4. W. B. Whitten and J. M. Ramsey, Anal. Chem. 63, 1027 (1991).
5. E. Betzig and R. J. Chichester, Science 262, 1422 (1993); J. J. Macklin, J. K. Trautman, T. D. Harris, L. E. Brus, ibid. 272, 255 (1996).
6. E. Betzig and R. J. Chichester, Science 262, 1422 (1993); J. J. Macklin, J. K. Trautman, T. D. Harris, L. E. Brus, ibid. 272, 255 (1996).
7. M. Eigen and R. Rigler, Proc. Natl. Acad. Sci. U.S.A. 91, 5740 (1994).
8. S. B. Smith, P. K. Aldridge, J. B. Callis, Science 243, 203 (1989); A. Castro, F. R. Fairfield, E. B. Shera, Anal. Chem. 65, 849 (1993); Y.-H. Lee, R. G. Maus, B. W. Smith, J. D. Winefordner, ibid. 66, 4142 (1994); M. D. Barnes, W. B. Whitten, J. M. Ramsey, ibid. 67, 418A (1995).
9. S. B. Smith, P. K. Aldridge, J. B. Callis, Science 243, 203 (1989); A. Castro, F. R. Fairfield, E. B. Shera, Anal. Chem. 65, 849 (1993); Y.-H. Lee, R. G. Maus, B. W. Smith, J. D. Winefordner, ibid. 66, 4142 (1994); M. D. Barnes, W. B. Whitten, J. M. Ramsey, ibid. 67, 418A (1995).
10. S. B. Smith, P. K. Aldridge, J. B. Callis, Science 243, 203 (1989); A. Castro, F. R. Fairfield, E. B. Shera, Anal. Chem. 65, 849 (1993); Y.-H. Lee, R. G. Maus, B. W. Smith, J. D. Winefordner, ibid. 66, 4142 (1994); M. D. Barnes, W. B. Whitten, J. M. Ramsey, ibid. 67, 418A (1995).
11. S. B. Smith, P. K. Aldridge, J. B. Callis, Science 243, 203 (1989); A. Castro, F. R. Fairfield, E. B. Shera, Anal. Chem. 65, 849 (1993); Y.-H. Lee, R. G. Maus, B. W. Smith, J. D. Winefordner, ibid. 66, 4142 (1994); M. D. Barnes, W. B. Whitten, J. M. Ramsey, ibid. 67, 418A (1995).
12. S. Nie, D. T. Chiu, R. N. Zare, Anal. Chem. 67, 2849 (1995); U. Mets and R. Rigler, J. Fluorescence 4, 259 (1994).
13. S. Nie, D. T. Chiu, R. N. Zare, Anal. Chem. 67, 2849 (1995); U. Mets and R. Rigler, J. Fluorescence 4, 259 (1994).
14. T. Funatsu, Y. Harada, M. Tokunaga, K. Saito, T. Yanagida, Nature 374, 555 (1995); R. D. Vale et al., ibid. 380, 451 (1996).
15. T. Funatsu, Y. Harada, M. Tokunaga, K. Saito, T. Yanagida, Nature 374, 555 (1995); R. D. Vale et al., ibid. 380, 451 (1996).
16. F.-R. F. Fan and A. J. Bard, Science 267, 871 (1995).
17. J. H. Jett et al., J. Biomol. Struct. Dyn. 7, 301 (1989).
18. M. M. Collinson and R. M. Wightman, Science 268, 1883 (1995).
19. Carboxy-rhodamine-6G (absorption maximum = 550 nm) was used to label a 30-base ssDNA to produce 5′-rhod-GAAAA, TGTGC, TGACC, GGACA, TG-AAA,ATGAG-3′ (DNA-R6G) with a molecular mass of 9773 g/mol. R6G (laser grade, ACROS, Trenton, NJ) and DNA-R6G (Baron Biotech, Milford, CT) were used without further purification. All solutions were prepared with deionized water from a Millipore Milli-Q system and then carefully filtered three times through a 0.22-μm filter (Costar, Cambridge, MA).
20. R. A. Mathies and K. Peck, Anal. Chem. 62, 1786 (1990); S. A. Soper et al., ibid. 63, 432 (1991); K. Ng, W. B. Whitten, S. Arnold, J. M. Ramsey, ibid. 64, 2914 (1992).
21. R. A. Mathies and K. Peck, Anal. Chem. 62, 1786 (1990); S. A. Soper et al., ibid. 63, 432 (1991); K. Ng, W. B. Whitten, S. Arnold, J. M. Ramsey, ibid. 64, 2914 (1992).
22. R. A. Mathies and K. Peck, Anal. Chem. 62, 1786 (1990); S. A. Soper et al., ibid. 63, 432 (1991); K. Ng, W. B. Whitten, S. Arnold, J. M. Ramsey, ibid. 64, 2914 (1992).
23. T. Schmidt, G. J. Schutz, W. Baumgartner, H. J. Gruber, H. Schindler, Proc. Natl. Acad. Sci. U.S.A. 93, 2926 (1996).
24. After submission of our work, a related paper appeared [R. M. Dickson, D. J. Norris, Y.-L. Tzeng, W. E. Moerner, Science 274, 966 (1996)]. By correlating the fluorescence spots in consecutive images, Dickson et al. followed molecular diffusional motion by constructing trajectories of motion. The images were recorded every 0.7 s, so only highly restricted motion, such as that found in certain pores of gel matrices, could be studied. Because evanescent-wave excitation was used, the observed intensities could be used to estimate the z position of the molecule, assuming there were no orientational or environmental effects on the spectroscopic properties. The molecules were not monitored continuously, so photobleaching could not be distinguished from movement away from the evanescent field. To study dynamics in free solution, as is demonstrated in our study, one needs a faster temporal response, the ability to observe the molecules continuously, and a way to decouple diffusion from photodecomposition.
25. 1/2 [L. B. Anderson and C. N. Reilley, J. Chem. Educ. 44, 9 (1967)].
26. -1) (7), based on the mass change, by use of the Stokes-Einstein equation [P. W. Atkins, Phys. Chem. (Freeman, San Francisco, CA, 1982), pp. 823-905].
27. J. Tellinghuisen, P. M. Goodwin, W. P. Ambrose, J. C. Martin, R. A. Keller, Anal. Chem. 66, 64 (1994).
28. J. Wang and P. Wolynes, Phys. Rev. Lett. 74, 4317 (1995).
29. The Ames Laboratory is operated for the U.S. Department of Energy by Iowa State University under contract W-7405-Eng-82. This work was supported by the Director of Energy Research, Office of Basic Energy Sciences, Division of Chemical Sciences.