OSA's Digital Library

Optics Express

Optics Express

  • Editor: Andrew M. Weiner
  • Vol. 22, Iss. 11 — Jun. 2, 2014
  • pp: 13710–13718

Three-axis digital holographic microscopy for high speed volumetric imaging

F. Saglimbeni, S. Bianchi, A. Lepore, and R. Di Leonardo  »View Author Affiliations

Optics Express, Vol. 22, Issue 11, pp. 13710-13718 (2014)

View Full Text Article

Enhanced HTML    Acrobat PDF (3610 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



Digital Holographic Microscopy allows to numerically retrieve three dimensional information encoded in a single 2D snapshot of the coherent superposition of a reference and a scattered beam. Since no mechanical scans are involved, holographic techniques have a superior performance in terms of achievable frame rates. Unfortunately, numerical reconstructions of scattered field by back-propagation leads to a poor axial resolution. Here we show that overlapping the three numerical reconstructions obtained by tilted red, green and blue beams results in a great improvement over the axial resolution and sectioning capabilities of holographic microscopy. A strong reduction in the coherent background noise is also observed when combining the volumetric reconstructions of the light fields at the three different wavelengths. We discuss the performance of our technique with two test objects: an array of four glass beads that are stacked along the optical axis and a freely diffusing rod shaped E.coli bacterium.

© 2014 Optical Society of America

OCIS Codes
(090.0090) Holography : Holography
(100.3010) Image processing : Image reconstruction techniques
(140.7010) Lasers and laser optics : Laser trapping
(180.0180) Microscopy : Microscopy
(180.6900) Microscopy : Three-dimensional microscopy
(090.1705) Holography : Color holography
(090.1995) Holography : Digital holography

ToC Category:

Original Manuscript: February 5, 2014
Revised Manuscript: March 25, 2014
Manuscript Accepted: March 28, 2014
Published: May 30, 2014

Virtual Issues
Vol. 9, Iss. 8 Virtual Journal for Biomedical Optics

F. Saglimbeni, S. Bianchi, A. Lepore, and R. Di Leonardo, "Three-axis digital holographic microscopy for high speed volumetric imaging," Opt. Express 22, 13710-13718 (2014)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. J. B. Pawley, Handbook of Biological Confocal Microscopy, 3 (Springer, 2006). [CrossRef]
  2. W. R. Zipfel, R. M. Williams, W. W. Webb, “Nonlinear magic: multiphoton microscopy in the biosciences,” Nat. Biotechnol. 21, 1369–1377 (2003). [CrossRef] [PubMed]
  3. P. Santi, “Light sheet fluorescence microscopy: a review,” J. Histochem. Cytochem. 59, 129–138 (2011). [CrossRef] [PubMed]
  4. K. B. Im, S. Han, H. Park, D. Kim, B. M. Kim, “Simple high-speed confocal line-scanning microscope,” Opt. Express 13, 5151–5156 (2005). [CrossRef] [PubMed]
  5. S. J. Lee, S. Kim, “Advanced particle-based velocimetry techniques for microscale flows,” Microfluid. Nanofluid. 6, 577–588 (2009).
  6. J. S. Dam, I. R. Perch-Nielsen, D. Palima, J. Glückstad, “Three-dimensional imaging in three-dimensional optical multi-beam micromanipulation,” Opt. Express 16, 7244–7250 (2008). [CrossRef] [PubMed]
  7. R. Bowman, G. Gibson, M. Padgett, “Particle tracking in stereomicroscopy in optical tweezers: Control of trap shape,” Opt. Express 18, 11785–11790 (2010). [CrossRef] [PubMed]
  8. M. P. Lee, G. M. Gibson, R. Bowman, S. Bernet, M. Ritsch-Marte, D. B. Phillips, M. J. Padgett, “A multi-modal stereo microscope based on a spatial light modulator,” Opt. Express 21, 16541–16551 (2013). [CrossRef] [PubMed]
  9. P. Memmolo, A. Finizio, M. Paturzo, L. Miccio, P. Ferraro, “Twin-beams digital holography for 3D tracking and quantitative phase-contrast microscopy in microfluidics,” Opt. Express 19, 25833–25842 (2011). [CrossRef]
  10. F. Charrière, A. Marian, F. Montfort, J. Kuehn, T. Colomb, E. Cuche, P. Marquet, C. Depeursinge, “Cell refractive index tomography by digital holographic microscopy,” Opt. Lett. 31, 178–180 (2006). [CrossRef] [PubMed]
  11. F. Charrière, N. Pavillon, T. Colomb, C. Depeursinge, T. J. Heger, E. A. D. Mitchell, P. Marquet, B. Rappaz, “Living specimen tomography by digital holographic microscopy: morphometry of testate amoeba,” Opt. Express 14, 7005–7013 (2006). [CrossRef] [PubMed]
  12. W. Choi, C. Fang-Yen, K. Badizadegan, S. Oh, N. Lue, R. R. Dasari, M. S. Feld, “Tomographic phase microscopy,” Nat. Methods 4, 717–719 (2007). [CrossRef] [PubMed]
  13. Y. Cotte, F. Toy, P. Jourdain, N. Pavillon, D. Boss, P. Magistretti, P. Marquet, C. Depeursinge, “Marker-free phase nanoscopy,” Nat. Photon. 7, 113–117 (2013). [CrossRef]
  14. K. Kim, K. S. Kim, H. Park, J. C. Ye, Y. Park, “Real-time visualization of 3-D dynamic microscopic objects using optical diffraction tomography,” Opt. Express 21, 32269–32278 (2013). [CrossRef]
  15. T. Kim, R. Zhou, M. Mir, S. D. Babacan, P. S. Carney, L. L. Goddard, G. Popescu, “White-light diffraction tomography of unlabeled live cells,” Nat. Photon. 8, 256–263 (2014). [CrossRef]
  16. M. K. Kim, Digital Holography and Microscopy: Principles, Techniques, and Applications (Springer, 2011). [CrossRef]
  17. W. Xu, M. H. Jericho, I. A. Meinertzhagen, H. J. Kreuzer, “Digital in-line holography of microspheres,” Appl. Opt. 41, 5367–5375 (2002). [CrossRef] [PubMed]
  18. J. Sheng, E. Malkiel, J. Katz, “Digital holographic microscope for measuring three-dimensional particle distributions and motions,” Appl. Opt. 45, 3893–3901 (2006). [CrossRef] [PubMed]
  19. Y. Park, G. Popescu, K. Badizadegan, R. R. Dasari, M. S. Feld, “Fresnel particle tracing in three dimensions using diffraction phase microscopy,” Opt. Lett. 32, 811–813 (2007). [CrossRef] [PubMed]
  20. L. Cavallini, G. Bolognesi, R. Di Leonardo, “Real-time digital holographic microscopy of multiple and arbitrarily oriented planes,” Opt. Lett. 36, 3491–3493 (2011). [CrossRef] [PubMed]
  21. J. Sheng, E. Malkiel, J. Katz, J. Adolf, R. Belas, A. R. Place, “Digital holographic microscopy reveals prey-induced changes in swimming behavior of predatory dinoflagellates,” Proc. Natl. Acad. Sci. U. S. A. 104, 17512–17517 (2007). [CrossRef] [PubMed]
  22. J. Garcia-Sucerquia, W. Xu, S. K. Jericho, P. Klages, M. H. Jericho, H. J. Kreuzer, “Digital in-line holographic microscopy,” Appl. Opt. 45, 836–850 (2006). [CrossRef] [PubMed]
  23. S. Lee, D. G. Grier, “Holographic microscopy of holographically trapped three-dimensional structures,” Opt. Express 15, 1505–1512 (2007). [CrossRef] [PubMed]
  24. S. Lee, Y. Roichman, G. Yi, S. Kim, S. Yang, A. van Blaaderen, P. van Oostrum, D. G. Grier, “Characterizing and tracking single colloidal particles with video holographic microscopy,” Opt. Express 15, 18275–18282 (2007). [CrossRef] [PubMed]
  25. F. Saglimbeni, S. Bianchi, G. Bolognesi, G. Paradossi, R. Di Leonardo, “Optical characterization of an individual polymer-shelled microbubble structure via digital holography,” Soft Matter 8, 8822–8825 (2012). [CrossRef]
  26. L. Repetto, E. Piano, C. Pontiggia, “Lensless digital holographic microscope with light-emitting diode illumination,” Opt. Lett. 29, 1132–1134 (2004). [CrossRef] [PubMed]
  27. F. Dubois, L. Joannes, J. Legros, “Improved three-dimensional imaging with a digital holography microscope with a source of partial spatial coherence,” Appl. Opt. 38, 7085–7094 (1999). [CrossRef]
  28. D. Tseng, O. Mudanyali, C. Oztoprak, S. O. Isikman, I. Sencan, O. Yagliderea, A. Ozcan, “Lensfree microscopy on a cellphone,” Lab Chip 10, 1787–1792 (2010). [CrossRef] [PubMed]
  29. M. Born, E. Wolf, Principles of Optics (Pergamon, 1993).
  30. F. Shen, A. Wang, “Fast-Fourier-transform based numerical integration method for the Rayleigh-Sommerfeld diffraction formula,” Appl. Opt. 45, 1102–1110 (2006). [CrossRef] [PubMed]
  31. S. Bianchi, R. Di Leonardo, “Real-time optical micro-manipulation using optimized holograms generated on the GPU,” Comput. Phys. Commun. 181, 1444–1448 (2010). [CrossRef]
  32. E. Cuche, P. Marquet, C. Depeursinge, “Simultaneous amplitude-contrast and quantitative phase-contrast microscopy by numerical reconstruction of Fresnel off-axis holograms,” Appl. Opt. 38, 6994–7001 (1999). [CrossRef]
  33. P. Marquet, B. Rappaz, P. J. Magistretti, E. Cuche, Y. Emery, T. Colomb, C. Depeursinge, “Digital holographic microscopy: a noninvasive contrast imaging technique allowing quantitative visualization of living cells with subwavelength axial accuracy,” Opt. Lett. 30, 468–470 (2005). [CrossRef] [PubMed]

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.


Fig. 1 Fig. 2 Fig. 3
Fig. 4 Fig. 5

Supplementary Material

» Media 1: MP4 (3192 KB)     

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited