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Biological applications of second harmonic imaging

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Abstract

Second Harmonic Generation (SHG) microscopy dates back to 1974, but effective biological use of the technique has a history of barely 10 years. It is now widely used to image collagen in many different applications, and is becoming useful for imaging myosin and some polysaccharides. A separate line on research has focussed on SHG dyes, which can provide high-speed indication of membrane potential and are now in use in neurobiology. This review looks at the progress to date in these different fields.

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References

  • Brüggemann DA, Brewer J, Risbo J, Bagatolli L (2010) Second harmonic generation microscopy: A tool for spatially and temporally resolved studies of heat induced structural changes in meat. Food Biophysics 5:1–8

    Article  Google Scholar 

  • Campagnola PJ, Clark HA, Mohler WA, Lewis A, Loew LM (2001) Second harmonic imaging microscopy of living cells. J Biomed Opt 6:277–286

    Article  PubMed  CAS  Google Scholar 

  • Campagnola PJ, Millard AC, Terasaki M, Hoppe PE, Malone CJ, Mohler WA (2002) Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues. Biophys J 81:493–508

    Article  Google Scholar 

  • Chen M-H, Chen WL, Sun Y, Fwu PT, Dong C-Y (2007) Multiphoton autofluorescence and second-harmonic generation imaging of the tooth. J Biomed Opt 12:064018

    Article  PubMed  Google Scholar 

  • Chu S-W, Chen I-H, Liu T-M, Chen PC, Sun CK (2001) Multimodal nonlinear spectral microscopy based on a femtosecond Cr:forsterite laser. Opt Lett 26:1909–1911

    Article  PubMed  CAS  Google Scholar 

  • Cox GC, Manconi F, Kable EPK (2002) Second harmonic imaging of collagen in mammalian tissue. Proc SPIE 4620:148–156

    Article  CAS  Google Scholar 

  • Cox GC, Kable E, Jones A, Fraser I, Manconi F, Gorrell M (2003) 3-dimensional imaging of collagen using second harmonic generation. J Struct Biol 141:53–62

    Article  PubMed  CAS  Google Scholar 

  • Cox GC, Kable EP (2006) Second harmonic imaging of collagen. In: Taatjes DJ, Mossman BT (eds) Cell imaging techniques. Methods in molecular biology, vol 319. Humana Press, Totowa, NJ, pp 15–35

    Chapter  Google Scholar 

  • Cox GC, Moreno N, Feijó JM (2005) Second-harmonic imaging of plant polysaccharides. J Biomed Opt 10:024013

    Article  PubMed  Google Scholar 

  • Denk W, Strickler JH, Webb WW (1990) Two photon laser scanning fluorescence microscopy. Science 248:73–76

    Article  PubMed  CAS  Google Scholar 

  • Dombeck DA, Sacconi L, Blanchard-Desce M, Webb WW (2005) Optical recording of fast neuronal membrane potential transients in acute mammalian brain slices by second-harmonic generation microscopy. J Neurophysiol 94:3628–3636

    Article  PubMed  Google Scholar 

  • Franken PA, Hill AE, Peters CW, Weinreich G (1961) Generation of optical harmonics. Phys Rev Lett 7:118–119

    Article  Google Scholar 

  • Freund I, Deutsch M, Sprecher A (1986) Connective tissue polarity. Optical second-harmonic microscopy, crossed-beam summation, and small-angle scattering in rat-tail tendon. Biophys J 50:693–712

    Article  PubMed  CAS  Google Scholar 

  • Fung DT, Sereysky JB, Basta-Pljakic J, Laudier DM, Huq R, Jepsen KJ, Schaffler MB, Flatow EL (2010) Second harmonic generation imaging and Fourier transform spectral analysis reveal damage in fatigue-loaded tendons. Ann Biomed Eng 38:1741–1751

    Article  PubMed  Google Scholar 

  • Gailhouste L, Le Grand Y, Odin C, Guyader D, Turlin B, Ezan F, Désille Y, Guilbert T, Bessard A, Frémin C, Theret N, Baffet G (2010) Fibrillar collagen scoring by second harmonic microscopy: a new tool in the assessment of liver fibrosis. J Hepatol 52:398–406

    Article  PubMed  CAS  Google Scholar 

  • Gannaway JN, Sheppard CJR (1978) Second harmonic imaging in the scanning optical microscope. Opt Quantum Electro 10:435–439

    Article  CAS  Google Scholar 

  • Georgiou E, Theodossiou T, Hovhannisya V, Politopoulos K, Rapti GS, Yova D (2000) Second and third optical harmonic generation in type I collagen, by nanosecond laser irradiation, over a broad spectral region. Opt Commun 176:253–260

    Article  CAS  Google Scholar 

  • Gorrell MD, Wang XM, Levy MT, Kable E, Marinos G, Cox G, McCaughan GW (2003) Intrahepatic expression of collagen and fibroblast activation protein (FAP) in hepatitis C virus infection. Adv Exp Med Biol 524:235–243

    Article  PubMed  CAS  Google Scholar 

  • Hellwarth R, Christensen P (1974) Nonlinear microscopic examination of structure in polycrystalline ZnSe. Opt Commun 12:318–322

    Article  CAS  Google Scholar 

  • Jiang J, Yuste R (2008) Second-harmonic generation imaging of membrane potential with photon counting. Microsc Microanal 14:526–531

    Article  PubMed  CAS  Google Scholar 

  • König K, Riemann I (2003) High-resolution multiphoton tomography of human skin with subcellular spatial resolution and picosecond time resolution. J Biomed Opt 8:432–439

    Article  PubMed  Google Scholar 

  • Lodish H, Berk A, Lipursky SL, Matsudaira P, Baltimore D, Darrell J (2000) Molecular cell biology, 4th edn. Freeman, New York

    Google Scholar 

  • Manconi F, Kable EP, Cox GC, Markham R, Fraser IS (2003) Whole-mount sections displaying microvascular and glandular structures in human uterus using multiphoton excitation microscopy. Micron 34:351–358

    Google Scholar 

  • Mizutani G, Sonoda Y, Sano H, Sakamoto M, Takahashi T, Ushioda S (2000) Detection of starch granules in a living plant by optical second harmonic microscopy. J Lumin 87:824–826

    Article  Google Scholar 

  • Mohler W, Millard AC, Campagnola PJ (2003) Second harmonic generation imaging of endogenous structural proteins. Methods 29:97–109

    Article  PubMed  CAS  Google Scholar 

  • Moreno N, Feijo J, Cox G (2004) Implementation and evaluation of a detector for forward propagated second harmonic signals. Micron 35:721–724

    Article  PubMed  Google Scholar 

  • Nadiarnykh O, Lacomb RB, Brewer MA, Campagnola PJ, et al (2010)Alterations of the extracellular matrix in ovarian cancer studied by Second Harmonic Generation imaging microscopy.BMC Cancer 10:94–142

    Google Scholar 

  • Nucciotti V, Stringari C, Sacconi L, Vanzi F, Fusi L, Linari M, Piazzesi G, Lombardi V, Pavone FS (2010) Probing myosin structural conformation in vivo bysecond-harmonic generation microscopy. Proc Natl Acad Sci USA 107:7763–7768

    Google Scholar 

  • Nuriya M, Jiang J, Nemet B, Eisenthal KB, Yuste R (2006) Imaging membrane potential in dendritic spines. Proc Nat Acad Sci USA 103:786–790

    Article  PubMed  CAS  Google Scholar 

  • Pons T, Moreaux L, Mongin O, Blanchard-Desce M, Mertz J (2003) Mechanisms of membrane potential sensing with second-harmonic generation microscopy. J Biomed Opt 8:428–431

    Article  PubMed  CAS  Google Scholar 

  • Provenzano PP, Eliceiri KW, Yan L, Ada-Nguema A, Conklin MW, Inman DR, Keely PJ (2008) Nonlinear optical imaging of cellular processes in breast cancer. Microsc Microanal 14:532–548

    Article  PubMed  CAS  Google Scholar 

  • Raja AM, Xu S, Zhou J, Tai DCS, Chen CS, Yu H (2010) Pulse-modulated second harmonic imaging microscope imaging quantitatively demonstrates marked increase of collagen in tumor after chemotherapy. J Biomed Opt 15:056016

    Article  PubMed  Google Scholar 

  • Ralston E, Swaim B, Czapiga M, Hwud W-L, Chien Y-H, Pittis MG, Bembi B, Schwartz O, Plotz P, Raben N (2008) Detection and imaging of non-contractile inclusions and sarcomeric anomalies in skeletal muscle by second harmonic generation combined with two-photon excited fluorescence. J Struct Biol 162:500–508

    Article  PubMed  CAS  Google Scholar 

  • Roth S, Freund I (1981) Optical second-harmonic scattering in rat-tail tendon. Biopolymers 20:1271–1290

    Article  PubMed  CAS  Google Scholar 

  • Sivaguru M, Durgam S, Ambekar R, Luedtke D, Fried G, Stewart A, Toussaint KC (2010) Quantitative analysis of collagen fiber organization in injured tendons using Fourier transform-second harmonic generation imaging. Opt Express 18:24983–24993

    Article  PubMed  CAS  Google Scholar 

  • Tai DC, Tan N, Xu S, Kang CH, Chia SM, Cheng CL, Wee A, Wei CL, Raja AM, Xiao G, Chang S, Rajapakse JC, So PT, Tang HH, Chen CS, Yu H (2009) Fibro-C-Index: comprehensive, morphology-based quantification of liver fibrosis using second harmonic generation and two-photon microscopy. J Biomed Opt 14(044013):1–10

    Google Scholar 

  • Wallace SJ, Morrison JL, Botting KJ, Kee TW (2008) Second-harmonic generation and two-photon-excited autofluorescence microscopy of cardiomyocytes: quantification of cell volume and myosin filaments. J Biomed Opt 13:064018

    Article  PubMed  Google Scholar 

  • Williams RM, Zipfel WR, Webb WW (2004) Interpreting second harmonic generation images of collagen I fibrils. Biophys J 88:1377–1386

    Article  PubMed  Google Scholar 

  • Xu P, Cox G, Ramshaw JAM, Lukins P and Sheppard CJR (2004) Polarization effects in SHG of collagen. Proceedings of SPIE 5323, 343–352

  • Xu P, Kable EP, Sheppard CJR, Cox G (2010) A quasi-crystal model of collagen microstructure based on SHG microscopy. Chin Opt Lett 8:213–216

    Article  CAS  Google Scholar 

  • Zipfel WR, Williams RM, Christie R, Nitikin AY, Hyman BT, Webb WW (2003) Live tissue intrinsic emission microscopy using multiphoton excited native fluorescence and second harmonic generation. Proc Nat Acad Sci USA 100:7075–7080

    Article  PubMed  CAS  Google Scholar 

  • Zhou R, Lu H, Liu X, Gong Y, Mao D (2010) Second-harmonic generation from a periodic array of noncentrosymmetric nanoholes. J Opt Soc Am B27:2405–2409

    Google Scholar 

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Correspondence to Guy Cox.

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Cox, G. Biological applications of second harmonic imaging. Biophys Rev 3, 131–141 (2011). https://doi.org/10.1007/s12551-011-0052-9

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