Pixel resolution polarization-sensitive second harmonic generation (PSHG) imaging has been recently shown as a promising imaging modality, by largely enhancing the capabilities of conventional intensity-based SHG microscopy. PSHG is able to obtain structural information from the elementary SHG active structures, which play an important role in many biological processes. Although the technique is of major interest, acquiring such information requires long offline processing, even with current computers. In this paper, we present an approach based on Fourier analysis of the anisotropy signature that allows processing the PSHG images in less than a second in standard single core computers. This represents a temporal improvement of several orders of magnitude compared to conventional fitting algorithms. This opens up the possibility for fast PSHG information with the subsequent benefit of potential use in medical applications.

© 2010 OSA

1. P. J. Campagnola, M. D. Wei, A. Lewis, and L. M. Loew, “High-resolution nonlinear optical imaging of live cells by second harmonic generation,” Biophys. J. 77(6), 3341–3349 (1999). [CrossRef] [PubMed]
2. M. E. Llewellyn, R. P. J. Barretto, S. L. Delp, and M. J. Schnitzer, “Minimally invasive high-speed imaging of sarcomere contractile dynamics in mice and humans,” Nature 454(7205), 784–788 (2008). [PubMed]
3. L. Fu and M. Gu, “Polarization anisotropy in fiber-optic second harmonic generation microscopy,” Opt. Express 16(7), 5000–5006 (2008). [CrossRef] [PubMed]
4. H. Bao, A. Boussioutas, R. Jeremy, S. Russell, and M. Gu, “Second harmonic generation imaging via nonlinear endomicroscopy,” Opt. Express 18(2), 1255–1260 (2010). [CrossRef] [PubMed]
5. E. U. Rafailov, M. A. Cataluna, and W. Sibbett, “Mode-locked quantum-dot lasers,” Nat. Photonics 1(7), 395–401 (2007). [CrossRef]
6. P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues,” Biophys. J. 82(1), 493–508 (2002). [CrossRef]
7. R. M. Williams, W. R. Zipfel, and W. W. Webb, “Interpreting second-harmonic generation images of collagen I fibrils,” Biophys. J. 88(2), 1377–1386 (2005). [CrossRef]
8. S.-W. Chu, S.-P. Tai, M.-C. Chan, C.-K. Sun, I.-C. Hsiao, C.-H. Lin, Y.-C. Chen, and B.-L. Lin, “Thickness dependence of optical second harmonic generation in collagen fibrils,” Opt. Express 15(19), 12005–12010 (2007). [CrossRef] [PubMed]
9. G. Recher, D. Rouède, P. Richard, A. Simon, J.-J. Bellanger, and F. Tiaho, “Three distinct sarcomeric patterns of skeletal muscle revealed by SHG and TPEF microscopy,” Opt. Express 17(22), 19763–19777 (2009). [CrossRef] [PubMed]
10. S. V. Plotnikov, A. M. Kenny, S. J. Walsh, B. Zubrowski, C. Joseph, V. L. Scranton, G. A. Kuchel, D. Dauser, M. Xu, C. C. Pilbeam, D. J. Adams, R. P. Dougherty, P. J. Campagnola, and W. A. Mohler, “Measurement of muscle disease by quantitative second-harmonic generation imaging,” J. Biomed. Opt. 13(4), 044018 (2008). [CrossRef] [PubMed]
11. O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12(5), 051805 (2007). [CrossRef] [PubMed]
12. S. Plotnikov, V. Juneja, A. B. Isaacson, W. A. Mohler, and P. J. Campagnola, “Optical clearing for improved contrast in second harmonic generation imaging of skeletal muscle,” Biophys. J. 90(1), 328–339 (2006). [CrossRef]
13. P. Matteini, F. Ratto, F. Rossi, R. Cicchi, C. Stringari, D. Kapsokalyvas, F. S. Pavone, and R. Pini, “Photothermally-induced disordered patterns of corneal collagen revealed by SHG imaging,” Opt. Express 17(6), 4868–4878 (2009). [CrossRef] [PubMed]
14. R. Cicchi, D. Kapsokalyvas, V. De Giorgi, V. Maio, A. Van Wiechen, D. Massi, T. Lotti, and F. S. Pavone, “Scoring of collagen organization in healthy and diseased human dermis by multiphoton microscopy,” J Biophoton. 3(1-2), 34–43 (2010). [CrossRef]
15. R. A. Rao, M. R. Mehta, and K. C. Toussaint., “Fourier transform-second-harmonic generation imaging of biological tissues,” Opt. Express 17(17), 14534–14542 (2009). [CrossRef] [PubMed]
16. R. A. R. Rao, M. R. Mehta, S. Leithem, and K. C. Toussaint., “Quantitative analysis of forward and backward second-harmonic images of collagen fibers using Fourier transform second-harmonic-generation microscopy,” Opt. Lett. 34(24), 3779–3781 (2009). [CrossRef] [PubMed]
17. K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12(6), 064019 (2007). [CrossRef]
18. P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82(6), 3330–3342 (2002). [CrossRef] [PubMed]
19. S. Plotnikov, V. Juneja, A. B. Isaacson, W. A. Mohler, and P. J. Campagnola, “Optical clearing for improved contrast in second harmonic generation imaging of skeletal muscle,” Biophys. J. 90(1), 328–339 (2006). [CrossRef]
20. F. Tiaho, G. Recher, and D. Rouède, “Estimation of helical angles of myosin and collagen by second harmonic generation imaging microscopy,” Opt. Express 15(19), 12286–12295 (2007). [CrossRef] [PubMed]
21. S. Psilodimitrakopoulos, S. I. Santos, I. Amat-Roldan, A. K. Thayil, D. Artigas, and P. Loza-Alvarez, “In vivo, pixel-resolution mapping of thick filaments’ orientation in nonfibrilar muscle using polarization-sensitive second harmonic generation microscopy,” J. Biomed. Opt. 14(1), 014001 (2009), http://spiedl.aip.org/getpdf/servlet/GetPDFServlet?filetype=pdf&id=JBOPFO000014000001014001000001&idtype=cvips&prog=normal . [CrossRef] [PubMed]
22. C. Odin, T. Guilbert, A. Alkilani, O. P. Boryskina, V. Fleury, and Y. Le Grand, “Collagen and myosin characterization by orientation field second harmonic microscopy,” Opt. Express 16(20), 16151–16165 (2008). [CrossRef] [PubMed]
23. S. Psilodimitrakopoulos, D. Artigas, G. Soria, I. Amat-Roldan, A. M. Planas, and P. Loza-Alvarez, “Quantitative discrimination between endogenous SHG sources in mammalian tissue, based on their polarization response,” Opt. Express 17(12), 10168–10176 (2009). [CrossRef] [PubMed]
24. W. L. Chen, T. H. Li, P. J. Su, C. K. Chou, P. T. Fwu, S. J. Lin, D. Kim, P. T. C. So, and C. Y. Dong, “Second harmonic generation chi tensor microscopy for tissue imaging,” Appl. Phys. Lett. 94, 3 (2009).
25. S. Psilodimitrakopoulos, V. Petegnief, G. Soria, I. Amat-Roldan, D. Artigas, A. M. Planas, and P. Loza-Alvarez, “Estimation of the effective orientation of the SHG source in primary cortical neurons,” Opt. Express 17(16), 14418–14425 (2009). [CrossRef] [PubMed]
26. S. Psilodimitrakopoulos, I. Amat-Roldan, P. Loza-Alvarez, and D. Artigas, “Estimating the helical pitch angle of amylopectin in starch using polarization second harmonic generation microscopy,” J. Opt. 12(8), 084007 (2010). [CrossRef]
27. S. Psilodimitrakopoulos, I. Amat-Roldan, S. Santos, M. Mathew, A.K.N. Thayil, D. Zalvidea, D. Artigas, and P. Loza-Alvarez, “Starch granules as a probe for the polarization at the sample plane of a high resolution multiphoton microscope,” SPIE, 68600E (2008).
28. O. Nadiarnykh and P. J. Campagnola, “Retention of polarization signatures in SHG microscopy of scattering tissues through optical clearing,” Opt. Express 17(7), 5794–5806 (2009). [CrossRef] [PubMed]

Appl. Phys. Lett.

• W. L. Chen, T. H. Li, P. J. Su, C. K. Chou, P. T. Fwu, S. J. Lin, D. Kim, P. T. C. So, and C. Y. Dong, “Second harmonic generation chi tensor microscopy for tissue imaging,” Appl. Phys. Lett. 94, 3 (2009).

Biophys. J.

• P. Stoller, K. M. Reiser, P. M. Celliers, and A. M. Rubenchik, “Polarization-modulated second harmonic generation in collagen,” Biophys. J. 82(6), 3330–3342 (2002). [CrossRef] [PubMed]
• S. Plotnikov, V. Juneja, A. B. Isaacson, W. A. Mohler, and P. J. Campagnola, “Optical clearing for improved contrast in second harmonic generation imaging of skeletal muscle,” Biophys. J. 90(1), 328–339 (2006). [CrossRef]
• P. J. Campagnola, M. D. Wei, A. Lewis, and L. M. Loew, “High-resolution nonlinear optical imaging of live cells by second harmonic generation,” Biophys. J. 77(6), 3341–3349 (1999). [CrossRef] [PubMed]
• P. J. Campagnola, A. C. Millard, M. Terasaki, P. E. Hoppe, C. J. Malone, and W. A. Mohler, “Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues,” Biophys. J. 82(1), 493–508 (2002). [CrossRef]
• R. M. Williams, W. R. Zipfel, and W. W. Webb, “Interpreting second-harmonic generation images of collagen I fibrils,” Biophys. J. 88(2), 1377–1386 (2005). [CrossRef]
• S. Plotnikov, V. Juneja, A. B. Isaacson, W. A. Mohler, and P. J. Campagnola, “Optical clearing for improved contrast in second harmonic generation imaging of skeletal muscle,” Biophys. J. 90(1), 328–339 (2006). [CrossRef]

J Biophoton.

• R. Cicchi, D. Kapsokalyvas, V. De Giorgi, V. Maio, A. Van Wiechen, D. Massi, T. Lotti, and F. S. Pavone, “Scoring of collagen organization in healthy and diseased human dermis by multiphoton microscopy,” J Biophoton. 3(1-2), 34–43 (2010). [CrossRef]

J. Biomed. Opt.

• K. M. Reiser, C. Bratton, D. R. Yankelevich, A. Knoesen, I. Rocha-Mendoza, and J. Lotz, “Quantitative analysis of structural disorder in intervertebral disks using second harmonic generation imaging: comparison with morphometric analysis,” J. Biomed. Opt. 12(6), 064019 (2007). [CrossRef]
• S. V. Plotnikov, A. M. Kenny, S. J. Walsh, B. Zubrowski, C. Joseph, V. L. Scranton, G. A. Kuchel, D. Dauser, M. Xu, C. C. Pilbeam, D. J. Adams, R. P. Dougherty, P. J. Campagnola, and W. A. Mohler, “Measurement of muscle disease by quantitative second-harmonic generation imaging,” J. Biomed. Opt. 13(4), 044018 (2008). [CrossRef] [PubMed]
• O. Nadiarnykh, S. Plotnikov, W. A. Mohler, I. Kalajzic, D. Redford-Badwal, and P. J. Campagnola, “Second harmonic generation imaging microscopy studies of osteogenesis imperfecta,” J. Biomed. Opt. 12(5), 051805 (2007). [CrossRef] [PubMed]
• S. Psilodimitrakopoulos, S. I. Santos, I. Amat-Roldan, A. K. Thayil, D. Artigas, and P. Loza-Alvarez, “In vivo, pixel-resolution mapping of thick filaments’ orientation in nonfibrilar muscle using polarization-sensitive second harmonic generation microscopy,” J. Biomed. Opt. 14(1), 014001 (2009), http://spiedl.aip.org/getpdf/servlet/GetPDFServlet?filetype=pdf&id=JBOPFO000014000001014001000001&idtype=cvips&prog=normal . [CrossRef] [PubMed]

J. Opt.

• S. Psilodimitrakopoulos, I. Amat-Roldan, P. Loza-Alvarez, and D. Artigas, “Estimating the helical pitch angle of amylopectin in starch using polarization second harmonic generation microscopy,” J. Opt. 12(8), 084007 (2010). [CrossRef]

Nat. Photonics

• E. U. Rafailov, M. A. Cataluna, and W. Sibbett, “Mode-locked quantum-dot lasers,” Nat. Photonics 1(7), 395–401 (2007). [CrossRef]

Nature

• M. E. Llewellyn, R. P. J. Barretto, S. L. Delp, and M. J. Schnitzer, “Minimally invasive high-speed imaging of sarcomere contractile dynamics in mice and humans,” Nature 454(7205), 784–788 (2008). [PubMed]

Opt. Express

• L. Fu and M. Gu, “Polarization anisotropy in fiber-optic second harmonic generation microscopy,” Opt. Express 16(7), 5000–5006 (2008). [CrossRef] [PubMed]
• H. Bao, A. Boussioutas, R. Jeremy, S. Russell, and M. Gu, “Second harmonic generation imaging via nonlinear endomicroscopy,” Opt. Express 18(2), 1255–1260 (2010). [CrossRef] [PubMed]
• S.-W. Chu, S.-P. Tai, M.-C. Chan, C.-K. Sun, I.-C. Hsiao, C.-H. Lin, Y.-C. Chen, and B.-L. Lin, “Thickness dependence of optical second harmonic generation in collagen fibrils,” Opt. Express 15(19), 12005–12010 (2007). [CrossRef] [PubMed]
• G. Recher, D. Rouède, P. Richard, A. Simon, J.-J. Bellanger, and F. Tiaho, “Three distinct sarcomeric patterns of skeletal muscle revealed by SHG and TPEF microscopy,” Opt. Express 17(22), 19763–19777 (2009). [CrossRef] [PubMed]
• R. A. Rao, M. R. Mehta, and K. C. Toussaint., “Fourier transform-second-harmonic generation imaging of biological tissues,” Opt. Express 17(17), 14534–14542 (2009). [CrossRef] [PubMed]
• P. Matteini, F. Ratto, F. Rossi, R. Cicchi, C. Stringari, D. Kapsokalyvas, F. S. Pavone, and R. Pini, “Photothermally-induced disordered patterns of corneal collagen revealed by SHG imaging,” Opt. Express 17(6), 4868–4878 (2009). [CrossRef] [PubMed]
• S. Psilodimitrakopoulos, V. Petegnief, G. Soria, I. Amat-Roldan, D. Artigas, A. M. Planas, and P. Loza-Alvarez, “Estimation of the effective orientation of the SHG source in primary cortical neurons,” Opt. Express 17(16), 14418–14425 (2009). [CrossRef] [PubMed]
• C. Odin, T. Guilbert, A. Alkilani, O. P. Boryskina, V. Fleury, and Y. Le Grand, “Collagen and myosin characterization by orientation field second harmonic microscopy,” Opt. Express 16(20), 16151–16165 (2008). [CrossRef] [PubMed]
• S. Psilodimitrakopoulos, D. Artigas, G. Soria, I. Amat-Roldan, A. M. Planas, and P. Loza-Alvarez, “Quantitative discrimination between endogenous SHG sources in mammalian tissue, based on their polarization response,” Opt. Express 17(12), 10168–10176 (2009). [CrossRef] [PubMed]
• F. Tiaho, G. Recher, and D. Rouède, “Estimation of helical angles of myosin and collagen by second harmonic generation imaging microscopy,” Opt. Express 15(19), 12286–12295 (2007). [CrossRef] [PubMed]
• O. Nadiarnykh and P. J. Campagnola, “Retention of polarization signatures in SHG microscopy of scattering tissues through optical clearing,” Opt. Express 17(7), 5794–5806 (2009). [CrossRef] [PubMed]

Opt. Lett.

• R. A. R. Rao, M. R. Mehta, S. Leithem, and K. C. Toussaint., “Quantitative analysis of forward and backward second-harmonic images of collagen fibers using Fourier transform second-harmonic-generation microscopy,” Opt. Lett. 34(24), 3779–3781 (2009). [CrossRef] [PubMed]

Other

• S. Psilodimitrakopoulos, I. Amat-Roldan, S. Santos, M. Mathew, A.K.N. Thayil, D. Zalvidea, D. Artigas, and P. Loza-Alvarez, “Starch granules as a probe for the polarization at the sample plane of a high resolution multiphoton microscope,” SPIE, 68600E (2008).

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