OSA's Digital Library

Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 18, Iss. 7 — Mar. 29, 2010
  • pp: 6563–6576

Precision analysis for standard deviation measurements of immobile single fluorescent molecule images

Michael C. DeSantis, Shawn H. DeCenzo, Je-Luen Li, and Y. M. Wang  »View Author Affiliations


Optics Express, Vol. 18, Issue 7, pp. 6563-6576 (2010)
http://dx.doi.org/10.1364/OE.18.006563


View Full Text Article

Enhanced HTML    Acrobat PDF (1149 KB)





Browse Journals / Lookup Meetings

Browse by Journal and Year


   


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools

Share
Citations

Abstract

Standard deviation measurements of intensity profiles of stationary single fluorescent molecules are useful for studying axial localization, molecular orientation, and a fluorescence imaging system’s spatial resolution. Here we report on the analysis of the precision of standard deviation measurements of intensity profiles of single fluorescent molecules imaged using an EMCCD camera. We have developed an analytical expression for the standard deviation measurement error of a single image which is a function of the total number of detected photons, the background photon noise, and the camera pixel size. The theoretical results agree well with the experimental, simulation, and numerical integration results. Using this expression, we show that single-molecule standard deviation measurements offer nanometer precision for a large range of experimental parameters.

© 2010 Optical Society of America

OCIS Codes
(100.6640) Image processing : Superresolution
(100.6890) Image processing : Three-dimensional image processing
(110.2960) Imaging systems : Image analysis
(180.2520) Microscopy : Fluorescence microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Image Processing

History
Original Manuscript: January 25, 2010
Revised Manuscript: March 1, 2010
Manuscript Accepted: March 2, 2010
Published: March 15, 2010

Virtual Issues
Vol. 5, Iss. 7 Virtual Journal for Biomedical Optics

Citation
Michael C. DeSantis, Shawn H. DeCenzo, Je-Luen Li, and Y. M. Wang, "Precision analysis for standard deviation measurements of immobile single fluorescent molecule images," Opt. Express 18, 6563-6576 (2010)
http://www.opticsinfobase.org/oe/abstract.cfm?URI=oe-18-7-6563


Sort:  Author  |  Year  |  Journal  |  Reset  

References

  1. R. E. Thompson, D. R. Larson, and W. W. Webb, "Precise nanometer localization analysis for individual fluorescent probes," Biophys. J. 82, 2775-2783 (2002). [CrossRef] [PubMed]
  2. A. Yildiz, J. N. Forkey, S. A. McKinney, T. Ha, Y. E. Goldman, and P. R. Selvin, "Myosin V walks hand-overhand: Single fluorophore imaging with 1.5-nm localization," Science 300, 2061-2065 (2003). [CrossRef] [PubMed]
  3. Y. M. Wang, R. H. Austin, and E. C. Cox, "Single molecule measurements of repressor protein 1D diffusion on DNA," Phys. Rev. Lett. 97, 048302 (2006). [CrossRef] [PubMed]
  4. C. Joo, H. Balci, Y. Ishitsuka, C. Buranachai, and T. Ha, "Advances in single-molecule fluorescence methods for molecular biology," Annu. Rev. Biochem. 77, 51-76 (2008). [CrossRef] [PubMed]
  5. A. M. van Oijen, J. Köhler, J. Schmidt, M. Müller, and G. J. Brakenhoff, "3-Dimensional super-resolution by spectrally selective imaging," Chem. Phys. Lett. 292, 183-187 (1998). [CrossRef]
  6. M. Speidel, A. Jonas, and E.-L. Florin, "Three-dimensional tracking of fluorescent nanoparticles with subnanometer precision by use of off-focus imaging," Opt. Lett. 28 (2003). [CrossRef] [PubMed]
  7. B. Huang, W. Wang, M. Bates, and X. Zhuang, "Three-dimensional super-resolution imaging by stochastic optical reconstruction microscopy," Science 319, 810-813 (2008). [CrossRef] [PubMed]
  8. K. Adachi, R. Yasuda, H. Noji, Y. Harada, M. Yoshida, and K. Kinosita, "Stepping rotation of F1-ATPase visualized through angle-resolved single-fluorophore imaging," Proc. Natl. Acad. Sci. USA 97, 7243-7247 (2000). [CrossRef] [PubMed]
  9. J. Enderlein, E. Toprak, and P. R. Selvin, "Polarization effect on position accuracy of fluorophore localization," Opt. Express 14, 8111-8120 (2006). [CrossRef] [PubMed]
  10. F. Aguet, S. Geissbühler, I. Märki, T. Lasser, and M. Unser, "Super-resolution orientation estimation and localization of fluorescent dipoles using 3-D steerable filters," Opt. Express 17, 6829-6848 (2009). [CrossRef] [PubMed]
  11. T. J. Holmes, D. Briggs, and A. A. Tarif, "Blind deconvolution," in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed. (Springer, New York, 2006), pp. 468-487. [CrossRef]
  12. A. Yildiz, M. Tomishige, R. D. Vale, and P. R. Selvin, "Kinesin walks hand-over-hand," Science 303, 676-678 (2004). [CrossRef]
  13. N. Bobroff, "Position measurement with a resolution and noise-limited instrument," Rev. Sci. Instrum. 57, 1152-1157 (1986). [CrossRef]
  14. M. H. Ulbrich and E. Y. Isacoff, "Subunit counting in membrane-bound proteins," Nature Methods 4, 319-321 (2007). [PubMed]
  15. J. Hynecek and T. Nishiwaki, "Excess noise and other important characteristics of low light level imaging using charge multiplying CCDs," IEEE Trans. on Electron Devices 50, 239-245 (2003). [CrossRef]
  16. J. R. Taylor, An Introduction to Error Analysis (University Science Books, California, 1997).
  17. Y. M. Wang, J. Tegenfeldt, W. Reisner, R. Riehn, X.-J. Guan, L. Guo, I. Golding, E. C. Cox, J. Sturm, and R. H. Austin, "Single-molecule studies of repressor-DNA interactions show long-range interactions," Proc. Natl. Acad. Sci. USA 102, 9796-9801 (2005). [CrossRef] [PubMed]
  18. M. Born and E. Wolf, Principles of Optics (Cambridge University Press, Cambridge, UK, 1999).
  19. S. H. DeCenzo, M. C. DeSantis, and Y. M. Wang, "Single-image measurements of unresolved dimolecular separations," In preparation (2010).
  20. S. K. G. Zareh, M. C. DeSantis, and Y. M. Wang, "Direct observation and analysis of 3D-diffusing fluorescent proteins in solution using single-image measurements," In preparation (2010).

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.

Figures

Fig. 1. Fig. 2. Fig. 3.
 

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited