OSA's Digital Library

Virtual Journal for Biomedical Optics

Virtual Journal for Biomedical Optics


  • Editors: Andrew Dunn and Anthony Durkin
  • Vol. 6, Iss. 9 — Oct. 3, 2011

Lensfree super-resolution holographic microscopy using wetting films on a chip

Onur Mudanyali, Waheb Bishara, and Aydogan Ozcan  »View Author Affiliations

Optics Express, Vol. 19, Issue 18, pp. 17378-17389 (2011)

View Full Text Article

Enhanced HTML    Acrobat PDF (2275 KB) Open Access

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



We investigate the use of wetting films to significantly improve the imaging performance of lensfree pixel super-resolution on-chip microscopy, achieving < 1 µm spatial resolution over a large imaging area of ~24 mm2. Formation of an ultra-thin wetting film over the specimen effectively creates a micro-lens effect over each object, which significantly improves the signal-to-noise-ratio and therefore the resolution of our lensfree images. We validate the performance of this approach through lensfree on-chip imaging of various objects having fine morphological features (with dimensions of e.g., ≤0.5 µm) such as Escherichia coli (E. coli), human sperm, Giardia lamblia trophozoites, polystyrene micro beads as well as red blood cells. These results are especially important for the development of highly sensitive field-portable microscopic analysis tools for resource limited settings.

© 2011 OSA

OCIS Codes
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(090.1995) Holography : Digital holography

ToC Category:

Original Manuscript: July 5, 2011
Revised Manuscript: July 30, 2011
Manuscript Accepted: August 3, 2011
Published: August 18, 2011

Virtual Issues
Vol. 6, Iss. 9 Virtual Journal for Biomedical Optics

Onur Mudanyali, Waheb Bishara, and Aydogan Ozcan, "Lensfree super-resolution holographic microscopy using wetting films on a chip," Opt. Express 19, 17378-17389 (2011)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. W. R. Zipfel, R. M. Williams, and W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21(11), 1369–1377 (2003). [CrossRef] [PubMed]
  2. S. W. Hell, “Toward fluorescence nanoscopy,” Nat. Biotechnol. 21(11), 1347–1355 (2003). [CrossRef] [PubMed]
  3. M. G. L. Gustafsson, “Nonlinear structured-illumination microscopy: wide-field fluorescence imaging with theoretically unlimited resolution,” Proc. Natl. Acad. Sci. U.S.A. 102(37), 13081–13086 (2005). [CrossRef] [PubMed]
  4. E. Betzig, G. H. Patterson, R. Sougrat, O. W. Lindwasser, S. Olenych, J. S. Bonifacino, M. W. Davidson, J. Lippincott-Schwartz, and H. F. Hess, “Imaging intracellular fluorescent proteins at nanometer resolution,” Science 313(5793), 1642–1645 (2006). [CrossRef] [PubMed]
  5. M. J. Rust, M. Bates, and X. Zhuang, “Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM),” Nat. Methods 3(10), 793–796 (2006). [CrossRef] [PubMed]
  6. S. T. Hess, T. P. K. Girirajan, and M. D. Mason, “Ultra-high resolution imaging by fluorescence photoactivation localization microscopy,” Biophys. J. 91(11), 4258–4272 (2006). [CrossRef] [PubMed]
  7. E. Chung, D. Kim, Y. Cui, Y.-H. Kim, and P. T. C. So, “Two-dimensional standing wave total internal reflection fluorescence microscopy: superresolution imaging of single molecular and biological specimens,” Biophys. J. 93(5), 1747–1757 (2007). [CrossRef] [PubMed]
  8. W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, and M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4(9), 717–719 (2007). [CrossRef] [PubMed]
  9. R. P. J. Barretto, B. Messerschmidt, and M. J. Schnitzer, “In vivo fluorescence imaging with high-resolution microlenses,” Nat. Methods 6(7), 511–512 (2009). [CrossRef] [PubMed]
  10. K. Goda, K. K. Tsia, and B. Jalali, “Serial time-encoded amplified imaging for real-time observation of fast dynamic phenomena,” Nature 458(7242), 1145–1149 (2009). [CrossRef] [PubMed]
  11. W. Hübner, G. P. McNerney, P. Chen, B. M. Dale, R. E. Gordon, F. Y. S. Chuang, X.-D. Li, D. M. Asmuth, T. Huser, and B. K. Chen, “Quantitative 3D video microscopy of HIV transfer across T cell virological synapses,” Science 323(5922), 1743–1747 (2009). [CrossRef] [PubMed]
  12. M. Bonnet, A. Ramsay, W. Githui, L. Gagnidze, F. Varaine, and P. J. Guerin, “Bleach sedimentation: an opportunity to optimize smear microscopy for tuberculosis diagnosis in settings of high prevalence of HIV,” Clin. Infect. Dis. 46(11), 1710–1716 (2008). [CrossRef] [PubMed]
  13. T. Nakada, H. Matsuzawa, H. Igarashi, Y. Fujii, and I. L. Kwee, “In vivo visualization of senile-plaque-like pathology in Alzheimer’s disease patients by MR microscopy on a 7T system,” J. Neuroimaging 18(2), 125–129 (2008). [CrossRef] [PubMed]
  14. D. V. Patel and C. N. J. McGhee, “In vivo confocal microscopy of corneal stromal nerves in patients with peripheral neuropathy,” Arch. Neurol. 66(9), 1179–1180, author reply 1180 (2009). [CrossRef] [PubMed]
  15. M. Selvarajah, K. Nicholls, T. D. Hewitson, and G. J. Becker, “Targeted urine microscopy in Anderson-Fabry Disease: a cheap, sensitive and specific diagnostic technique,” Nephrol. Dial. Transplant. (2011). doi:10.1093/ndt/gfr084.
  16. T. Kojima, Y. Matsumoto, M. Dogru, and K. Tsubota, “The application of in vivo laser scanning confocal microscopy as a tool of conjunctival in vivo cytology in the diagnosis of dry eye ocular surface disease,” Mol. Vis. 16, 2457–2464 (2010). [PubMed]
  17. A. Sartori, A. Mallet, E. Veiga, M. Bonazzi, S. Mostowy, L. Dortet, W. Baumeister, and P. Cossart, “Correlative Light/Electron Microscopy: a Tool for Investigating Infectious Diseases,” MAM 15(S2), 862 (2009). [CrossRef]
  18. Y. R. Shea, J. L. Davis, L. Huang, J. A. Kovacs, H. Masur, F. Mulindwa, S. Opus, Y. Chow, and P. R. Murray, “High sensitivity and specificity of acid-fast microscopy for diagnosis of pulmonary tuberculosis in an African population with a high prevalence of human immunodeficiency virus,” J. Clin. Microbiol. 47(5), 1553–1555 (2009). [CrossRef] [PubMed]
  19. S.-R. Wu, R. Loving, B. Lindqvist, H. Hebert, P. J. B. Koeck, M. Sjoberg, and H. Garoff, “Single-particle cryoelectron microscopy analysis reveals the HIV-1 spike as a tripod structure,” Proc. Natl. Acad. Sci. U.S.A. 107(44), 18844–18849 (2010). [CrossRef] [PubMed]
  20. B. J. Marais, W. Brittle, K. Painczyk, A. C. Hesseling, N. Beyers, E. Wasserman, D. van Soolingen, and R. M. Warren, “Use of light-emitting diode fluorescence microscopy to detect acid-fast bacilli in sputum,” Clin. Infect. Dis. 47(2), 203–207 (2008). [CrossRef] [PubMed]
  21. D. C. Essaka, J. White, P. Rathod, C. D. Whitmore, O. Hindsgaul, M. M. Palcic, and N. J. Dovichi, “Monitoring the uptake of glycosphingolipids in Plasmodium falciparum-infected erythrocytes using both fluorescence microscopy and capillary electrophoresis with laser-induced fluorescence detection,” Anal. Chem. 82(23), 9955–9958 (2010). [CrossRef] [PubMed]
  22. M. Krause, P. Rosch, B. Radt, and J. Popp, “Localizing and identifying living bacteria in an abiotic environment by a combination of Raman and fluorescence microscopy,” Anal. Chem. 80(22), 8568–8575 (2008). [CrossRef] [PubMed]
  23. S. E.-D. Hassan, S. I. Okoued, M. A. Mudathir, and E. M. Malik, “Testing the sensitivity and specificity of the fluorescence microscope (Cyscope) for malaria diagnosis,” Malar. J. 9(1), 88 (2010). [CrossRef] [PubMed]
  24. W. Bishara, U. Sikora, O. Mudanyali, T.-W. Su, O. Yaglidere, S. Luckhart, and A. Ozcan, “Holographic pixel super-resolution in portable lensless on-chip microscopy using a fiber-optic array,” Lab Chip 11(7), 1276–1279 (2011). [CrossRef] [PubMed]
  25. W. Bishara, T.-W. Su, A. F. Coskun, and A. Ozcan, “Lensfree on-chip microscopy over a wide field-of-view using pixel super-resolution,” Opt. Express 18(11), 11181–11191 (2010). [CrossRef] [PubMed]
  26. W. Bishara, H. Zhu, and A. Ozcan, “Holographic opto-fluidic microscopy,” Opt. Express 18(26), 27499–27510 (2010). [CrossRef] [PubMed]
  27. S. O. Isikman, W. Bishara, S. Mavandadi, F. W. Yu, S. Feng, R. Lau, and A. Ozcan, “Lens-free optical tomographic microscope with a large imaging volume on a chip,” Proc. Natl. Acad. Sci. U.S.A. 108(18), 7296–7301 (2011). [CrossRef] [PubMed]
  28. E. Cuche, F. Bevilacqua, and C. Depeursinge, “Digital holography for quantitative phase-contrast imaging,” Opt. Lett. 24(5), 291–293 (1999). [CrossRef] [PubMed]
  29. F. Dubois, L. Joannes, and J. C. Legros, “Improved three-dimensional imaging with a digital holography microscope with a source of partial spatial coherence,” Appl. Opt. 38(34), 7085–7094 (1999). [CrossRef] [PubMed]
  30. W. Xu, M. H. Jericho, I. A. Meinertzhagen, and H. J. Kreuzer, “Digital in-line holography for biological applications,” Proc. Natl. Acad. Sci. U.S.A. 98(20), 11301–11305 (2001). [CrossRef] [PubMed]
  31. G. Pedrini and H. J. Tiziani, “Short-coherence digital microscopy by use of a lensless holographic imaging system,” Appl. Opt. 41(22), 4489–4496 (2002). [CrossRef] [PubMed]
  32. L. Repetto, E. Piano, and C. Pontiggia, “Lensless digital holographic microscope with light-emitting diode illumination,” Opt. Lett. 29(10), 1132–1134 (2004). [CrossRef] [PubMed]
  33. T.-C. Poon, “Recent progress in optical scanning holography,” J. Hologr. Speckle 1(1), 6–25 (2004). [CrossRef]
  34. B. Javidi, I. Moon, S. K. Yeom, and E. Carapezza, “Three-dimensional imaging and recognition of microorganism using single-exposure on-line (SEOL) digital holography,” Opt. Express 13(12), 4492–4506 (2005). [CrossRef] [PubMed]
  35. C. J. Mann, L. F. Yu, C. M. Lo, and M. K. Kim, “High-resolution quantitative phase-contrast microscopy by digital holography,” Opt. Express 13(22), 8693–8698 (2005). [CrossRef] [PubMed]
  36. J. Rosen, G. Indebetouw, and G. Brooker, “Homodyne scanning holography,” Opt. Express 14(10), 4280–4285 (2006). [CrossRef] [PubMed]
  37. J. Garcia-Sucerquia, W. Xu, M. H. Jericho, and H. J. Kreuzer, “Immersion digital in-line holographic microscopy,” Opt. Lett. 31(9), 1211–1213 (2006). [CrossRef] [PubMed]
  38. G. Popescu, T. Ikeda, R. R. Dasari, and M. S. Feld, “Diffraction phase microscopy for quantifying cell structure and dynamics,” Opt. Lett. 31(6), 775–777 (2006). [CrossRef] [PubMed]
  39. F. Dubois, C. Yourassowsky, O. Monnom, J.-C. Legros, O. Debeir, P. Van Ham, R. Kiss, and C. Decaestecker, “Digital holographic microscopy for the three-dimensional dynamic analysis of in vitro cancer cell migration,” J. Biomed. Opt. 11(5), 054032 (2006). [CrossRef] [PubMed]
  40. A. Stern and B. Javidi, “Theoretical analysis of three-dimensional imaging and recognition of microorganisms using single-exposure on-line (SEOL) holographic microscope,” J. Opt. Soc. Am. A 24(1), 163–168 (2007). [CrossRef]
  41. S. S. Kou and C. J. R. Sheppard, “Imaging in digital holographic microscopy,” Opt. Express 15(21), 13640–13648 (2007). [CrossRef] [PubMed]
  42. J. Rosen and G. Brooker, “Non-scanning motionless fluorescence three-dimensional holographic microscopy,” Nat. Photonics 2(3), 190–195 (2008). [CrossRef]
  43. U. Gopinathan, G. Pedrini, and W. Osten, “Coherence effects in digital in-line holographic microscopy,” J. Opt. Soc. Am. A 25(10), 2459–2466 (2008). [CrossRef] [PubMed]
  44. V. Mico, Z. Zalevsky, C. Ferreira, and J. García, “Superresolution digital holographic microscopy for three-dimensional samples,” Opt. Express 16(23), 19260–19270 (2008). [CrossRef] [PubMed]
  45. M. Paturzo, F. Merola, S. Grilli, S. De Nicola, A. Finizio, and P. Ferraro, “Super-resolution in digital holography by a two-dimensional dynamic phase grating,” Opt. Express 16(21), 17107–17118 (2008). [CrossRef] [PubMed]
  46. N. T. Shaked and J. Rosen, “Multiple-viewpoint projection holograms synthesized by spatially incoherent correlation with broadband functions,” J. Opt. Soc. Am. A 25(8), 2129–2138 (2008). [CrossRef] [PubMed]
  47. D. J. Brady, K. Choi, D. L. Marks, R. Horisaki, and S. Lim, “Compressive holography,” Opt. Express 17(15), 13040–13049 (2009). [CrossRef] [PubMed]
  48. T. R. Hillman, T. Gutzler, S. A. Alexandrov, and D. D. Sampson, “High-resolution, wide-field object reconstruction with synthetic aperture Fourier holographic optical microscopy,” Opt. Express 17(10), 7873–7892 (2009). [CrossRef] [PubMed]
  49. L. Waller, Y. Luo, S. Y. Yang, and G. Barbastathis, “Transport of intensity phase imaging in a volume holographic microscope,” Opt. Lett. 35(17), 2961–2963 (2010). [CrossRef] [PubMed]
  50. N. T. Shaked, T. M. Newpher, M. D. Ehlers, and A. Wax, “Parallel on-axis holographic phase microscopy of biological cells and unicellular microorganism dynamics,” Appl. Opt. 49(15), 2872–2878 (2010). [CrossRef] [PubMed]
  51. D. Bonn, J. Eggers, J. Indekeu, J. Meunier, and E. Rolley, “Wetting and spreading,” Rev. Mod. Phys. 81(2), 739–805 (2009). [CrossRef]
  52. P. de Gennes, “Wetting: statics and dynamics,” Rev. Mod. Phys. 57(3), 827–863 (1985). [CrossRef]
  53. V. M. Starov, M. G. Velarde, and C. J. Radke, Wetting and Spreading Dynamics (CRC Press, 2007).
  54. N. A. Stelmashenko, J. P. Craven, A. M. Donald, E. M. Terentjev, and B. L. Thiel, “Topographic contrast of partially wetting water droplets in environmental scanning electron microscopy,” J. Microsc. 204(2), 172–183 (2001). [CrossRef] [PubMed]
  55. Q. F. Wei, R. R. Mather, A. F. Fotheringham, and R. D. Yang, “Dynamic Wetting of Fibers Observed in an Environmental Scanning Electron Microscope,” Text. Res. J. 73(6), 557–561 (2003). [CrossRef]
  56. E. Bormashenko, Y. Bormashenko, T. Stein, G. Whyman, R. Pogreb, and Z. Barkay, “Environmental scanning electron microscopy study of the fine structure of the triple line and cassie-wenzel wetting transition for sessile drops deposited on rough polymer substrates,” Langmuir 23(8), 4378–4382 (2007). [CrossRef] [PubMed]
  57. C. P. Allier, G. Hiernard, V. Poher, and J. M. Dinten, “Bacteria detection with thin wetting film lensless imaging,” Biomed. Opt. Express 1(3), 762–770 (2010). [CrossRef] [PubMed]
  58. C. P. Allier, V. Poher, J. G. Coutard, G. Hiernard, and J. M. Dinten, “Thin wetting film lensless imaging,” in Proceedings of the SPIE (2011), pp. 790608–790608–8.
  59. O. Mudanyali, D. Tseng, C. Oh, S. O. Isikman, I. Sencan, W. Bishara, C. Oztoprak, S. Seo, B. Khademhosseini, and A. Ozcan, “Compact, light-weight and cost-effective microscope based on lensless incoherent holography for telemedicine applications,” Lab Chip 10(11), 1417–1428 (2010). [CrossRef] [PubMed]
  60. S. Seo, S. O. Isikman, I. Sencan, O. Mudanyali, T.-W. Su, W. Bishara, A. Erlinger, and A. Ozcan, “High-throughput lens-free blood analysis on a chip,” Anal. Chem. 82(11), 4621–4627 (2010). [CrossRef] [PubMed]
  61. O. Mudanyali, C. Oztoprak, D. Tseng, A. Erlinger, and A. Ozcan, “Detection of waterborne parasites using field-portable and cost-effective lensfree microscopy,” Lab Chip 10(18), 2419–2423 (2010). [CrossRef] [PubMed]
  62. T.-W. Su, A. Erlinger, D. Tseng, and A. Ozcan, “Compact and light-weight automated semen analysis platform using lensfree on-chip microscopy,” Anal. Chem. 82(19), 8307–8312 (2010). [CrossRef] [PubMed]
  63. D. Tseng, O. Mudanyali, C. Oztoprak, S. O. Isikman, I. Sencan, O. Yaglidere, and A. Ozcan, “Lensfree microscopy on a cellphone,” Lab Chip 10(14), 1787–1792 (2010). [CrossRef] [PubMed]
  64. G. Stybayeva, O. Mudanyali, S. Seo, J. Silangcruz, M. Macal, E. Ramanculov, S. Dandekar, A. Erlinger, A. Ozcan, and A. Revzin, “Lensfree holographic imaging of antibody microarrays for high-throughput detection of leukocyte numbers and function,” Anal. Chem. 82(9), 3736–3744 (2010). [CrossRef] [PubMed]
  65. R. C. Hardie, K. J. Barnard, and E. E. Armstrong, “Joint MAP registration and high-resolution image estimation using a sequence of undersampled images,” IEEE Trans. Image Process. 6(12), 1621–1633 (1997). [CrossRef] [PubMed]
  66. S. C. Park, M. K. Park, and M. G. Kang, “Super-resolution image reconstruction: a technical overview,” IEEE Signal Process. Mag. 20(3), 21–36 (2003). [CrossRef]
  67. N. A. Woods, N. P. Galatsanos, and A. K. Katsaggelos, “Stochastic methods for joint registration, restoration, and interpolation of multiple undersampled images,” IEEE Trans. Image Process. 15(1), 201–213 (2006). [CrossRef] [PubMed]
  68. R. C. Hardie, “High-resolution image reconstruction from a sequence of rotated and translated frames and its application to an infrared imaging system,” Opt. Eng. 37(1), 247 (1998). [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