OSA's Digital Library

Optics Express

Optics Express

  • Editor: C. Martijn de Sterke
  • Vol. 16, Iss. 25 — Dec. 8, 2008
  • pp: 20891–20901

Polarization distortion effects in polarimetric two-photon microscopy

Peter Schön, Fabiana Munhoz, Alicja Gasecka, Sophie Brustlein, and Sophie Brasselet  »View Author Affiliations

Optics Express, Vol. 16, Issue 25, pp. 20891-20901 (2008)

View Full Text Article

Enhanced HTML    Acrobat PDF (379 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



We present a global analysis of experimental factors affecting polarization responses in two-photon inverted microscopy. The role of reflection optics and high numerical aperture focusing is investigated in two-photon fluorescence, which can be extended to other nonlinear processes. We show that both effects strongly distort polarization responses and can lead to misleading extraction of molecular order information from polarimetric measurements. We describe a model accounting for these effects and develop a calibration technique for the determination of polarization parameters in the sample plane using two-photon fluorescence polarimetry in liquids.

© 2008 Optical Society of America

OCIS Codes
(120.5410) Instrumentation, measurement, and metrology : Polarimetry
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:

Original Manuscript: August 13, 2008
Revised Manuscript: September 23, 2008
Manuscript Accepted: September 24, 2008
Published: December 3, 2008

Virtual Issues
Vol. 4, Iss. 2 Virtual Journal for Biomedical Optics

Peter Schön, Fabiana Munhoz, Alicja Gasecka, Sophie Brustlein, and Sophie Brasselet, "Polarization distortion effects in polarimetric two-photon microscopy," Opt. Express 16, 20891-20901 (2008)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. J. A. Dix and A. S. Verkman, "Mapping of fluorescence anisotropy in living cells by ratio imaging," Biophys. J. 57, 231-240 (1990).
  2. T. H. Foster, B. D. Pearson, S. Mitra, and C. E. Bigelow, "Fluorescence anisotropy imaging reveals localization of meso-tetrahydroxyphenyl chlorin in the nuclear envelope," Photochem. Photobiol. 81, 1544-1547 (2005). [CrossRef] [PubMed]
  3. J. Borejdo and S. Burlacu, "Measuring Orientation of Actin Filaments within a Cell: orientation of Actin in Intestinal Microvilli," Biophys. J. 65300-309 (1993). [CrossRef] [PubMed]
  4. R. K. P. Benninger, Björn önfelt, M. A. A. Neil, D. M. Davis, and P. M. W. French, "Fluorescence Imaging of Two-Photon Linear Dichroism: Cholesterol Depletion Disrupts Molecular Orientation in Cell Membranes," Biophys. J. 88609-622 (2005). [CrossRef]
  5. R. E. Dale, S. C. Hopkins,U. A. an der Heide, T. Marszałek, M. Irving, and Y. E. Goldman, "Model-Independent Analysis of the Orientation of Fluorescent Probes with Restricted Mobility in Muscle Fibers," Biophys. J. 781606-1618 (1999). [CrossRef]
  6. A. M. Vrabioiu and T. J. Mitchison, "Structural insights into yeast septin organization from polarized fluorescence microscopy," Nature 443, 466-468 (2006). [CrossRef] [PubMed]
  7. T. E. Schaus, E. W. Taylor, and G. G. Borisy, "Self-organization of actin filament orientation in the dendritic-nucleation/array-treadmilling model," Proc. Nat. Acad. Sc. 104, 7086-7091 (2007). [CrossRef]
  8. D. A. Dombeck, K. A. Kasischke, H. D. Vishwasrao, M. Ingelsson, B. T. Hyman, and W. W. Webb, "Uniform polarity microtubule assemblies imaged in native brain tissue by second harmonic generation microscopy," Proc. Nat. Acad. Sc. 100, 7081-7086 (2003). [CrossRef]
  9. D. Débarre, W. Supatto, A. M. Pena, A. Fabre, T. Tordjmann, L. Combettes, M. C. Schanne-Klein, and E. Beaurepaire, " Imaging lipid bodies in cells and tissues using third harmonic generation microscopy," Nature Methods 3, 47-53 (2006). [CrossRef]
  10. A. M. Pena, T. Boulesteix, T. Dartigalongue, M. Strupler, E. Beaurepaire, and M. C. Schanne-Klein, "Chiroptical effects in the second harmonic generation from collagens I and IV: applications in nonlinear microscopy," Nonlinear Opt. Quantum Opt. 35 (2006).
  11. S. Brasselet and J. Zyss, "Nonlinear polarimetry of molecular crystals down to the nanoscale," C. R. Phys. 8, 165-179 (2007). [CrossRef]
  12. C. Odin, Y. Le Grand, A. Renault, L. Gailhouste, and G. Baffet, "Orientation fields of nonlinear biological fibrils by second harmonic generation microscopy," J. Microsc. 229, 32-38 (2008). [CrossRef] [PubMed]
  13. V. Le Floc’h, S. Brasselet, J. F. Roch, and J. Zyss, "Monitoring of orientation in molecular ensembles by polarization sensitive nonlinear microscopy," J. Phys. Chem. B 107, 12403-12410 (2003). [CrossRef]
  14. C. Anceau, S. Brasselet, and J. Zyss, "Local orientational distribution of molecular monolayers probed by nonlinear microscopy," Chem. Phys. Lett. 411, 98-102 (2005). [CrossRef]
  15. S. Brasselet, V. Le Floc’h, F. Treussart, J. F. Roch, J. Zyss, E. Botzung-Appert, and A. Ibanez, "In situ diagnostics of the crystalline nature of single organic nanocrystals by nonlinear microscopy," Phys. Rev. Lett. 92, 207401 (2004). [CrossRef] [PubMed]
  16. D. Oron, D. Yelin, E. Tal, S. Raz, R. Fachima, and Y. Silberberg, "Depth-resolved structural imaging by third-harmonic generation microscopy," J. Struct. Biol. 147, 3-11 (2004). [CrossRef] [PubMed]
  17. H. Wang, Y. Fu, P. Zickmund, R. Shi, and J.-X. Cheng, "Coherent Anti-Stokes Raman Scattering Imaging of Axonal Myelin in Live Spinal Tissues," Biophys. J. 89, 581-591 (2005). [CrossRef] [PubMed]
  18. D. Lara and C. Dainty, "Axially resolved complete Mueller matrix confocal microscopy," Appl. Opt. 45, 1917-1930 (2006). [CrossRef] [PubMed]
  19. C. E. Bigelow and T.H. Foster, "Confocal fluorescence polarization microscopy in turbid media: Effects of scattering-induced depolarisation," J. Opt. Soc. Am. A 23, 2932 (2006). [CrossRef]
  20. T. Brixner, G. Krampert, P. Niklaus, and G. Gerber, "Generation and characterization of polarization-shaped femtosecond laser pulses," Appl. Phys. B 74, S133-S144 (2002). [CrossRef]
  21. D. Oron, N. Dudovich, and Y. Silberberg, "Femtosecond Phase-and-Polarization Control for Background-Free Coherent Anti-Stokes Raman Spectroscopy," Phys. Rev. Lett. 90, 213902 (2003). [CrossRef] [PubMed]
  22. E. Y. S. Yew, and ColinJ. R. Sheppard, "Effects of axial field components on second harmonic generation microscopy," Opt. Express 14, 1167-1174 (2006). [CrossRef] [PubMed]
  23. B. Richards and E. Wolf, "Electromagnetic diffraction in optical systems. II. Structure of the image field in an aplanetic system," Proc. R. Soc. London Ser. A. 153, 358-579 (1959).
  24. L. Novotny and B. Hecht, Principles of Nano-Optics (Cambridge University Press, Cambridge 2006).
  25. D. Axelrod, "Carbocyanine dye orientation in red cell membrane studied by microscopic fluorescence polarization," Biophys. J. 26, 557-574 (1979). [CrossRef] [PubMed]
  26. K. Komorowska, S. Brasselet, J. Zyss, L. Pourlsen, M. Jazdzyk, H. J. Egelhaaf, J. Gierschner, and M. Hanack, " Nanometric scale investigation of the nonlinear efficiency of perhydrotriphynylene inclusion compounds," Chem. Phys. 318, 12-20 (2005). [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