OSA's Digital Library

Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 20, Iss. 20 — Sep. 24, 2012
  • pp: 22245–22251

A novel method of diffraction imaging flow cytometry for sizing microspheres

Sa Yu, Jun Zhang, Marina S. Moran, Jun Q. Lu, Yuanming Feng, and Xin-Hua Hu  »View Author Affiliations

Optics Express, Vol. 20, Issue 20, pp. 22245-22251 (2012)

View Full Text Article

Enhanced HTML    Acrobat PDF (1039 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



We report a novel method of diffraction imaging flow cytometry to measure and analyze size distribution of microspheres. An automated and robust image processing software based on the short-time-Fourier-transform algorithm has been developed to analyze the characteristic and spatially varying oscillations of side scatters recorded as a diffraction image. Our results demonstrate that the new method allows accurate and rapid determination of single microspheres’ diameters ranging from 1 to 100μm. The capacity for analysis of light scattering by two-sphere aggregates has been demonstrated but analytical tools for characterization of aggregates by multiple microspheres remain to be developed.

© 2012 OSA

OCIS Codes
(110.1650) Imaging systems : Coherence imaging
(290.5850) Scattering : Scattering, particles

ToC Category:
Imaging Systems

Original Manuscript: June 29, 2012
Revised Manuscript: August 23, 2012
Manuscript Accepted: August 24, 2012
Published: September 13, 2012

Sa Yu, Jun Zhang, Marina S. Moran, Jun Q. Lu, Yuanming Feng, and Xin-Hua Hu, "A novel method of diffraction imaging flow cytometry for sizing microspheres," Opt. Express 20, 22245-22251 (2012)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. K. K. Kim, D. W. Pack, M. Ferrari, A. P. Lee, and L. J. Lee, “Microspheres for drug delivery,” in Biological and Biomedical Nanotechnology, A. P. Lee, J. Lee, and M. Ferrari, eds. (Springer, 2006), pp. 19–50.
  2. D. Svoboda, M. Kozubek, and S. Stejskal, “Generation of digital phantoms of cell nuclei and simulation of image formation in 3D image cytometry,” Cytometry A 75A(6), 494–509 (2009). [CrossRef] [PubMed]
  3. B. P. Hanley, L. Xing, and R. H. Cheng, “Variance in multiplex suspension array assays: microsphere size variation impact,” Theor. Biol. Med. Model. 4(1), 31–38 (2007). [CrossRef] [PubMed]
  4. I. K. Ludlow and P. H. Kaye, “A scanning diffractometer for the rapid analysis of microparticles and biological cells,” J. Colloid Interface Sci. 69(3), 571–589 (1979). [CrossRef]
  5. S. L. Min and A. Gomez, “High-resolution size measurement of single spherical particles with a fast Fourier transform of the angular scattering intensity,” Appl. Opt. 35(24), 4919–4926 (1996). [CrossRef] [PubMed]
  6. J. T. Soini, A. V. Chernyshev, P. E. Hänninen, E. Soini, and V. P. Maltsev, “A new design of the flow cuvette and optical set-up for the scanning flow cytometer,” Cytometry 31(2), 78–84 (1998). [CrossRef] [PubMed]
  7. K. A. Semyanov, P. A. Tarasov, A. E. Zharinov, A. V. Chernyshev, A. G. Hoekstra, and V. P. Maltsev, “Single-particle sizing from light scattering by spectral decomposition,” Appl. Opt. 43(26), 5110–5115 (2004). [CrossRef] [PubMed]
  8. C. Godefroy and M. Adjouadi, “Particle sizing in a flow environment using light scattering patterns,” Part. Part. Syst. Charact. 17(2), 47–55 (2000). [CrossRef]
  9. B. Ovryn, “Three-dimensional forward scattering particle image velocimetry applied to a microscopic field-of-view,” Exp. Fluids 29(7), S175–S184 (2000). [CrossRef]
  10. Y. L. Xu and B. A. S. Gustafson, “Comparison between Multisphere Light-scattering Calculations: Rigorous Solution and Discrete-Dipole Approximation,” Astrophys. J. 513(2), 894–909 (1999). [CrossRef]
  11. K. M. Jacobs, J. Q. Lu, and X. H. Hu, “Development of a diffraction imaging flow cytometer,” Opt. Lett. 34(19), 2985–2987 (2009). [CrossRef] [PubMed]
  12. K. M. Jacobs, L. V. Yang, J. Ding, A. E. Ekpenyong, R. Castellone, J. Q. Lu, and X. H. Hu, “Diffraction imaging of spheres and melanoma cells with a microscope objective,” J Biophotonics 2(8-9), 521–527 (2009). [CrossRef] [PubMed]
  13. K. Dong, Y. Feng, K. M. Jacobs, J. Q. Lu, R. S. Brock, L. V. Yang, F. E. Bertrand, M. A. Farwell, and X. H. Hu, “Label-free classification of cultured cells through diffraction imaging,” Biomed. Opt. Express 2(6), 1717–1726 (2011). [CrossRef] [PubMed]
  14. M. Portnoff, “Time-frequency representation of digital signals and systems based on short-time Fourier analysis,” IEEE Trans. Acoust., Speech Signal Process. 28(1), 55–69 (1980). [CrossRef]
  15. J. Q. Lu, P. Yang, and X.-H. Hu, “Simulations of light scattering from a biconcave red blood cell using the finite-difference time-domain method,” J. Biomed. Opt. 10(2), 024022 (2005). [CrossRef] [PubMed]
  16. M. A. Yurkin, A. G. Hoekstra, R. S. Brock, and J. Q. Lu, “Systematic comparison of the discrete dipole approximation and the finite difference time domain method for large dielectric scatterers,” Opt. Express 15(26), 17902–17911 (2007). [CrossRef] [PubMed]
  17. X. Ma, J. Q. Lu, R. S. Brock, K. M. Jacobs, P. Yang, and X. H. Hu, “Determination of complex refractive index of polystyrene microspheres from 370 to 1610 nm,” Phys. Med. Biol. 48(24), 4165–4172 (2003). [CrossRef] [PubMed]
  18. J. G. Daugman, “Complete discrete 2-D Gabor transforms by neural networks for image analysis and compression,” IEEE Trans. Acoust., Speech, Signal Process., 36(7), 1169–1179 (1988). [CrossRef]

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