Biological Second and Third Harmonic Generation Microscopy

Multiphoton microscopy has become a standard method for noninvasive imaging of thick specimens with subcellular resolution. Higher harmonic generation microscopy (HHGM), based on nonlinear multiphoton excitation, is a contrast mechanism for the structural and molecular imaging of native samples in cell culture and in fixed and live tissues, for both, three‐dimensional and four‐dimensional reconstructions. HHGM comprises second and third harmonic generation (SHG, THG) of ordered molecules, can be obtained without exogenous labels, and provides detailed real‐time optical reconstruction of fibrillar collagen, myosin, microtubules, and membrane potential, as well as cell depolarization. This unit presents the principles of SHG and THG and the basic setup of a HHGM system, and summarizes current applications in cell biology. Multimodal multiphoton microscopy using HHGM together with two‐photon excited fluorescence will develop into a key approach to real‐time imaging of cell dynamics in the context of live tissues.

[1]  Bernard Choi,et al.  Reversible dissociation of collagen in tissues. , 2003, The Journal of investigative dermatology.

[2]  Yaron Silberberg,et al.  Depth-resolved structural imaging by third-harmonic generation microscopy. , 2004, Journal of structural biology.

[3]  Watt W Webb,et al.  Interpreting second-harmonic generation images of collagen I fibrils. , 2005, Biophysical journal.

[4]  A. Fabre,et al.  Imaging lipid bodies in cells and tissues using third-harmonic generation microscopy , 2005, Nature Methods.

[5]  Takeshi Yasui,et al.  Characterization of collagen orientation in human dermis by two-dimensional second-harmonic-generation polarimetry. , 2004, Journal of biomedical optics.

[6]  Martin Vogel,et al.  Second harmonic imaging of intrinsic signals in muscle fibers in situ. , 2004, Journal of biomedical optics.

[7]  Leonardo Sacconi,et al.  Optical recording of fast neuronal membrane potential transients in acute mammalian brain slices by second-harmonic generation microscopy. , 2005, Journal of neurophysiology.

[8]  F S Pavone,et al.  Second-harmonic generation sensitivity to transmembrane potential in normal and tumor cells. , 2005, Journal of biomedical optics.

[9]  R Gauderon,et al.  Simultaneous multichannel nonlinear imaging: combined two-photon excited fluorescence and second-harmonic generation microscopy , 2000, SPIE Photonics Taiwan.

[10]  Jerome Mertz,et al.  Mechanisms of membrane potential sensing with second-harmonic generation microscopy. , 2003, Journal of biomedical optics.

[11]  S. Chu,et al.  Nonlinear bio‐photonic crystal effects revealed with multimodal nonlinear microscopy , 2002, Journal of microscopy.

[12]  B. Bellhouse,et al.  Identification of second harmonic optical effects from vaccine coated gold microparticles. , 2004, Physics in medicine and biology.

[13]  Guy Cox,et al.  3-dimensional imaging of collagen using second harmonic generation. , 2003, Journal of structural biology.

[14]  Watt W. Webb,et al.  Uniform polarity microtubule assemblies imaged in native brain tissue by second-harmonic generation microscopy , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[15]  E. Bröcker,et al.  Reconstructing leukocyte migration in 3D extracellular matrix by time-lapse videomicroscopy and computer-assisted tracking. , 2004, Methods in molecular biology.

[16]  Bruce J Tromberg,et al.  Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy. , 2004, Biophysical journal.

[17]  Daniel A Dombeck,et al.  Optical Recording of Action Potentials with Second-Harmonic Generation Microscopy , 2004, The Journal of Neuroscience.

[18]  Tsung-Han Tsai,et al.  Studies of chi(2)/chi(3) tensors in submicron-scaled bio-tissues by polarization harmonics optical microscopy. , 2004, Biophysical journal.

[19]  William A Mohler,et al.  Three-dimensional high-resolution second-harmonic generation imaging of endogenous structural proteins in biological tissues. , 2002, Biophysical journal.

[20]  Guy Cox,et al.  Second-harmonic imaging of plant polysaccharides. , 2005, Journal of biomedical optics.

[21]  K. König,et al.  Multiphoton autofluorescence imaging of intratissue elastic fibers. , 2005, Biomaterials.

[22]  K. König,et al.  Multiphoton microscopy in life sciences , 2000, Journal of microscopy.

[23]  R D Schaller,et al.  Nonlinear chemical imaging microscopy: near-field third harmonic generation imaging of human red blood cells. , 2000, Analytical chemistry.

[24]  J. Zavadil,et al.  Single cell behavior in metastatic primary mammary tumors correlated with gene expression patterns revealed by molecular profiling. , 2002, Cancer research.

[25]  W. Denk,et al.  Two-photon laser scanning fluorescence microscopy. , 1990, Science.

[26]  Michel Orrit,et al.  Third-harmonic generation from single gold nanoparticles. , 2005, Nano letters.

[27]  Peter Friedl,et al.  Compensation mechanism in tumor cell migration , 2003, The Journal of cell biology.

[28]  Chi-Kuang Sun,et al.  Higher harmonic generation microscopy. , 2005, Advances in biochemical engineering/biotechnology.

[29]  Tsung-Han Tsai,et al.  Studies of χ(2)/χ(3) Tensors in Submicron-Scaled Bio-Tissues by Polarization Harmonics Optical Microscopy , 2004 .

[30]  J. Mora,et al.  In vivo imaging of leukocyte trafficking in blood vessels and tissues. , 2004, Current opinion in immunology.

[31]  P. Friedl.  Dynamic imaging of cellular interactions with extracellular matrix , 2004, Histochemistry and Cell Biology.

[32]  W. Webb,et al.  Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[33]  Beop-Min Kim,et al.  Polarization-dependent optical second-harmonic imaging of a rat-tail tendon. , 2002, Journal of biomedical optics.

[34]  Leslie M Loew,et al.  Sensitivity of second harmonic generation from styryl dyes to transmembrane potential. , 2004, Biophysical journal.

[35]  Fu-Jen Kao,et al.  The use of optical parametric oscillator for harmonic generation and two‐photon UV fluorescence microscopy , 2004, Microscopy research and technique.

[36]  Paul J Campagnola,et al.  Second harmonic generation imaging of endogenous structural proteins. , 2003, Methods.

[37]  P. Friedl,et al.  Migration of highly aggressive MV3 melanoma cells in 3-dimensional collagen lattices results in local matrix reorganization and shedding of alpha2 and beta1 integrins and CD44. , 1997, Cancer research.

[38]  B. Lawn,et al.  Fracture of ceramic/ceramic/polymer trilayers for biomechanical applications. , 2003, Journal of biomedical materials research. Part A.

[39]  William A Mohler,et al.  Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres. , 2006, Biophysical journal.

[40]  S. Henrickson,et al.  T-cell priming by dendritic cells in lymph nodes occurs in three distinct phases , 2004, Nature.

[41]  A. Abbott.  Cell culture: Biology's new dimension , 2003, Nature.

[42]  A. Pena,et al.  Micrometer scale Ex Vivo multiphoton imaging of unstained arterial wall structure , 2006, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[43]  Chen-Yuan Dong,et al.  Characterizing the thermally induced structural changes to intact porcine eye, part 1: second harmonic generation imaging of cornea stroma. , 2005, Journal of biomedical optics.