Nonlinear Microscopy of the Vocal Folds

  • Mathias Strupler
  • Romain Deterre
  • Nadir Goulamhoussen
  • Fouzi Benboujja
  • Christopher J. Hartnick
  • Caroline Boudoux
Chapter
First Online:

Abstract

Nonlinear microscopy is becoming a very important tool available to life scientists. This powerful three-dimensional technique allows exploration of unstained biological tissues through a contrast provided by the nonlinear interaction of short laser pulses with certain macromolecules such as elastin and collagen. The possibility of imaging microstructures (cells, nuclei) as well as macromolecules without affecting the integrity of the organ paves the way for a better understanding of vocal folds’ normal and pathological conditions. In this chapter, we review the physical concepts behind nonlinear microscopy and provide example of its use in laryngology.

Keywords

Nonlinear microscopy Collagen Elastin Vocal folds Intrinsic contrast High-resolution imaging 
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Notes

Acknowledgments

The authors would like to thank Dr Amber Beckley, Mr. Étienne de Montigny, Ms. Chloé Gariépy, Mr. Scott Infusino, Pr. Steven Maturo, and Dr. Shilpa Ojha for fruitful discussions. Pr. Boudoux acknowledges funding from the National Sciences and Engineering Research Council of Canada (NSERC) and the Canadian Foundation for Innovation (CFI).

References

  1. 1.
    Huang D, Swanson E, Lin C, et al. Optical coherence tomography. Science. 1991;254(5035):1178–81. doi: 10.1126/science.1957169.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Ridgway JM, Armstrong WB, Guo S, et al. In vivo optical coherence tomography of the human oral cavity and oropharynx. Arch Otolaryngol Head Neck Surg. 2006;132(10):1074–81. doi: 10.1001/archotol.132.10.1074.CrossRefPubMedGoogle Scholar
  3. 3.
    Bouma BE, Tearney GJ, Compton CC, Nishioka NS. High-resolution imaging of the human esophagus and stomach in vivo using optical coherence tomography. Gastrointest Endosc. 2000;51(4):467–74. doi: 10.1016/S0016-5107(00)70449-4.CrossRefPubMedGoogle Scholar
  4. 4.
    Bus MTJ, Muller BG, de Bruin DM, et al. Volumetric in vivo visualization of upper urinary tract tumors using optical coherence tomography: a pilot study. J Urol. 2013;190(6):2236–42. doi: 10.1016/j.juro.2013.08.006.CrossRefPubMedGoogle Scholar
  5. 5.
    Yabushita H, Bouma BE, Houser SL, et al. Characterization of human atherosclerosis by optical coherence tomography. Circulation. 2002;106(13):1640–5. Available at: http://www.ncbi.nlm.nih.gov/pubmed/12270856. Accessed 12 Mar 2014.CrossRefPubMedGoogle Scholar
  6. 6.
    Hanna N, Saltzman D, Mukai D, et al. Two-dimensional and 3-dimensional optical coherence tomographic imaging of the airway, lung, and pleura. J Thorac Cardiovasc Surg. 2005;129(3):615–22. doi: 10.1016/j.jtcvs.2004.10.022.CrossRefPubMedGoogle Scholar
  7. 7.
    Wong BJF, Jackson RP, Guo S, et al. In vivo optical coherence tomography of the human larynx: normative and benign pathology in 82 patients. Laryngoscope. 2005;115(11):1904–11. doi: 10.1097/01.MLG.0000181465.17744.BE.CrossRefPubMedGoogle Scholar
  8. 8.
    Kraft M, Glanz H, von Gerlach S, Wisweh H, Lubatschowski H, Arens C. Clinical value of optical coherence tomography in laryngology. Head Neck. 2008;30(12):1628–35. doi: 10.1002/hed.20914.CrossRefPubMedGoogle Scholar
  9. 9.
    Armstrong WB, Ridgway JM, Vokes DE, et al. Optical coherence tomography of laryngeal cancer. Laryngoscope. 2006;116(7):1107–13. doi: 10.1097/01.mlg.0000217539.27432.5a.CrossRefPubMedGoogle Scholar
  10. 10.
    Burns JA, Zeitels SM, Anderson RR, Kobler JB, Pierce MC, de Boer JF. Imaging the mucosa of the human vocal fold with optical coherence tomography. Ann Otol Rhinol Laryngol. 2005;114(9):671–6. Available at: http://www.ncbi.nlm.nih.gov/pubmed/16240928. Accessed 12 Mar 2014.CrossRefPubMedGoogle Scholar
  11. 11.
    Klein A, Pierce M. Imaging the human vocal folds in vivo with optical coherence tomography: a preliminary experience. Ann Otol Rhinol Laryngol. 2006;115(4):277–84.CrossRefPubMedGoogle Scholar
  12. 12.
    Boudoux C, Leuin SC, Oh WY, et al. Preliminary evaluation of noninvasive microscopic imaging techniques for the study of vocal fold development. J Voice. 2009;23(3):269–76. Available at: http://dx.doi.org/10.1016/j.jvoice.2007.10.003.CrossRefPubMedGoogle Scholar
  13. 13.
    Boudoux C, Leuin SC, Oh WY, et al. Optical microscopy of the pediatric vocal fold. Arch Otolaryngol Head Neck Surg. 2009;135(1):53–64. Available at: http://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=19153308&retmode=ref&cmd=prlinks.CrossRefPubMedGoogle Scholar
  14. 14.
    Maturo S, Benboujja F, Boudoux C, Hartnick C. Quantitative distinction of unique vocal fold subepithelial architectures using optical coherence tomography. Ann Otol Rhinol Laryngol. 2012;121(11):754–60. Available at: http://www.ncbi.nlm.nih.gov/pubmed/23193909. Accessed 12 Mar 2014.CrossRefPubMedGoogle Scholar
  15. 15.
    Benboujja F, Rogers D, Infusino S, Strupler M, Hartnick CJ, Boudoux C. A study of vocal fold maturation using optical coherence tomography. San Francisco, CA: SPIE Photonics West; 2014.Google Scholar
  16. 16.
    Ridgway JM, Su J, Wright R, et al. Optical coherence tomography of the newborn airway. Ann Otol Rhinol Laryngol. 2008;117(5):327–34. Available at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2871770&tool=pmcentrez&rendertype=abstract. Accessed 12 Mar 2014.PubMedPubMedCentralGoogle Scholar
  17. 17.
    Wisweh H, Merkel U. Optical coherence tomography monitoring of vocal fold femtosecond laser microsurgery. European Conference on Biomedical Optics. Munich, Germany; 2007Google Scholar
  18. 18.
    Minsky M. Microscopy apparatus. 1961. Available at: http://www.google.ca/patents/US3013467. Accessed 4 Dec 2013.
  19. 19.
    Pawley J. Handbook of biological confocal microscopy. New York: Springer; 2006.CrossRefGoogle Scholar
  20. 20.
    Webb RH, Hughes GW, Delori FC. Confocal scanning laser ophthalmoscope. Appl Opt. 1987;26(8):1492–9. doi: 10.1364/AO.26.001492.CrossRefPubMedGoogle Scholar
  21. 21.
    Gareau DS, Jeon H, Nehal KS, Rajadhyaksha M. Rapid screening of cancer margins in tissue with multimodal confocal microscopy. J Surg Res. 2012;178(2):533–8. doi: 10.1016/j.jss.2012.05.059.CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Hsiung P-L, Hsiung P-L, Hardy J, et al. Detection of colonic dysplasia in vivo using a targeted heptapeptide and confocal microendoscopy. Nat Med. 2008;14(4):454–8. doi: 10.1038/nm1692.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
  24. 24.
    Boudoux C, Yun S-H, Oh W, et al. Rapid wavelength-swept spectrally encoded confocal microscopy. Opt Express. 2005;13(20):8214–21.CrossRefPubMedGoogle Scholar
  25. 25.
    Boudoux C, Benboujja F, Deterre R, Strupler M, Maturo S, Hartnick CJ. Emerging microscopy techniques for pediatric vocal fold evaluation. In: Izdebski K, Yan Y, Ward R, Wong BJF, editors. Normal & Abnormal Vocal Folds Kinematics: High-Speed Digital Phonoscopy (HSDP), Optical Coherence Tomography (OCT) & Narrow Band Imaging (NBI). San Francisco, CA: Pacific Vo.; 2014.Google Scholar
  26. 26.
    Iftimia N, Ferguson RD, Mujat M, et al. Combined reflectance confocal microscopy/optical coherence tomography imaging for skin burn assessment. Biomed Opt Express. 2013;4(5):680–95. doi: 10.1364/BOE.4.000680.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Kang D, Suter MJ, Boudoux C, et al. Co-registered spectrally encoded confocal microscopy and optical frequency domain imaging system. J Microsc. 2010;239(2):87–91. Available at:  http://doi.wiley.com/10.1111/j.1365-2818.2010.03367.x.
  28. 28.
    Zipfel WR, Williams RM, Webb WW. Nonlinear magic: multiphoton microscopy in the biosciences. Nat Biotechnol. 2003;21(11):1369–77. doi: 10.1038/nbt899.CrossRefPubMedGoogle Scholar
  29. 29.
    Rivera DR, Brown CM, Ouzounov DG, et al. Compact and flexible raster scanning multiphoton endoscope capable of imaging unstained tissue. Proc Natl Acad Sci U S A. 2011;108(43):17598–603. doi: 10.1073/pnas.1114746108.CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Engelbrecht CJ, Johnston RS, Seibel EJ, Helmchen F. Ultra-compact fiber-optic two-photon microscope for functional fluorescence imaging in vivo. Opt Express. 2008;16(8):5556–64. Available at: http://www.ncbi.nlm.nih.gov/pubmed/18542658. Accessed 18 April 2014.CrossRefPubMedGoogle Scholar
  31. 31.
    Helmchen F, Fee MS, Tank DW, Denk W. A miniature head-mounted two-photon microscope. High-resolution brain imaging in freely moving animals. Neuron. 2001;31(6):903–12. Available at: http://www.ncbi.nlm.nih.gov/pubmed/11580892. Accessed 31 Mar 2014.CrossRefPubMedGoogle Scholar
  32. 32.
    Miri AK, Tripathy U, Mongeau L, Wiseman PW. Nonlinear laser scanning microscopy of human vocal folds. Laryngoscope. 2012;122(2):356–63. doi: 10.1002/lary.22460.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Deterre R. Microscopie non-linéaire pour l’imagerie des cordes vocales. 2012.Google Scholar
  34. 34.
    Hoy CL, Everett WN, Yildirim M, Kobler J, Zeitels SM, Ben-Yakar A. Towards endoscopic ultrafast laser microsurgery of vocal folds. J Biomed Opt. 2012;17(3):038002. doi: 10.1117/1.JBO.17.3.038002.CrossRefPubMedGoogle Scholar
  35. 35.
    Boyd RW. Nonlinear optics. San Diego, CA: Acad. Press; 2003. p. 578. Available at: http://books.google.ca/books/about/Nonlinear_Optics.html?id=30t9VmOmOGsC&pgis=1. Accessed 9 Jul 2014.Google Scholar
  36. 36.
    Denk W, Strickler J, Webb W. Two-photon laser scanning fluorescence microscopy. Science. 1990;248(4951):73–6. doi: 10.1126/science.2321027.CrossRefPubMedGoogle Scholar
  37. 37.
    French PMW, Williams JAR, Taylor JR. Femtosecond pulse generation from a titanium-doped sapphire laser using nonlinear external cavity feedback. Opt Lett. 1989;14(13):686. doi: 10.1364/OL.14.000686.CrossRefPubMedGoogle Scholar
  38. 38.
    Denk W. Two-photon scanning photochemical microscopy: mapping ligand-gated ion channel distributions. Proc Natl Acad Sci U S A. 1994;91(14):6629–33. Available at: http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=44256&tool=pmcentrez&rendertype=abstract. Accessed 4 Dec 2013.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Brown EB, Campbell RB, Tsuzuki Y, et al. In vivo measurement of gene expression, angiogenesis and physiological function in tumors using multiphoton laser scanning microscopy. Nat Med. 2001;7(7):864–8. doi: 10.1038/89997.CrossRefPubMedGoogle Scholar
  40. 40.
    Sipkins DA, Wei X, Wu JW, et al. In vivo imaging of specialized bone marrow endothelial microdomains for tumour engraftment. Nature. 2005;435(7044):969–73. doi: 10.1038/nature03703.CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Squirrell JM, Wokosin DL, White JG, Bavister BD. Long-term two-photon fluorescence imaging of mammalian embryos without compromising viability. Nat Biotechnol. 1999;17(8):763–7. doi: 10.1038/11698.CrossRefPubMedGoogle Scholar
  42. 42.
    Sandberg LB, Soskel NT, Leslie JG. Elastin structure, biosynthesis, and relation to disease states. N Engl J Med. 1981;304:566–79.CrossRefPubMedGoogle Scholar
  43. 43.
    Debelle L, Tamburro AM. Elastin: molecular description and function. Int J Biochem Cell Biol. 1999;31(2):261–72. doi: 10.1016/S1357-2725(98)00098-3.CrossRefPubMedGoogle Scholar
  44. 44.
    Gray SD, Titze IR, Alipour F, Hammond TH. Biomechanical and histologic observations of vocal fold fibrous proteins. Ann Otol Rhinol Laryngol. 2000;109(1):77–85. Available at: http://europepmc.org/abstract/MED/10651418. Accessed 9 Jul 2014.CrossRefPubMedGoogle Scholar
  45. 45.
    Moore J, Thibeault S. Insights into the role of elastin in vocal fold health and disease. J Voice. 2012;26(3):269–75. doi: 10.1016/j.jvoice.2011.05.003.CrossRefPubMedGoogle Scholar
  46. 46.
    Breunig HG, Studier H, König K. Multiphoton excitation characteristics of cellular fluorophores of human skin in vivo. Opt Express. 2010;18(8):7857–71. Available at: http://www.ncbi.nlm.nih.gov/pubmed/20588627. Accessed 9 Jul 2014.CrossRefPubMedGoogle Scholar
  47. 47.
    Boulesteix T, Pena A-M, Pagès N, et al. Micrometer scale ex vivo multiphoton imaging of unstained arterial wall structure. Cytometry A. 2006;69(1):20–6. doi: 10.1002/cyto.a.20196.CrossRefPubMedGoogle Scholar
  48. 48.
    Zipfel WR, Williams RM, Christie R, Nikitin AY, Hyman BT, Webb WW. Live tissue intrinsic emission microscopy using multiphoton-excited native fluorescence and second harmonic generation. Proc Natl Acad Sci U S A. 2003;100(12):7075–80. doi: 10.1073/pnas.0832308100.CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Kierdaszuk B, Malak H, Gryczynski I, Callis P, Lakowicz JR. Fluorescence of reduced nicotinamides using one- and two-photon excitation. Biophys Chem. 1996;62(1–3):1–13. Available at: http://www.ncbi.nlm.nih.gov/pubmed/8962467. Accessed 10 Jul 2014.CrossRefPubMedGoogle Scholar
  50. 50.
    Huang S, Heikal AA, Webb WW. Two-photon fluorescence spectroscopy and microscopy of NAD(P)H and flavoprotein. Biophys J. 2002;82(5):2811–25. doi: 10.1016/S0006-3495(02)75621-X.CrossRefPubMedPubMedCentralGoogle Scholar
  51. 51.
    Strupler M, Pena A-M, Hernest M, et al. Second harmonic imaging and scoring of collagen in fibrotic tissues. Opt Express. 2007;15(7):4054–65. Available at: http://www.ncbi.nlm.nih.gov/pubmed/19532649. Accessed 9 Jul 2014.CrossRefPubMedGoogle Scholar
  52. 52.
    Pena A-M, Boulesteix T, Dartigalongue T, Schanne-Klein M-C. Chiroptical effects in the second harmonic signal of collagens I and IV. J Am Chem Soc. 2005;127(29):10314–22. doi: 10.1021/ja0520969.CrossRefPubMedGoogle Scholar
  53. 53.
    Deniset-Besseau A, Duboisset J, Benichou E, Hache F, Brevet P-F, Schanne-Klein M-C. Measurement of the second-order hyperpolarizability of the collagen triple helix and determination of its physical origin. J Phys Chem B. 2009;113(40):13437–45. doi: 10.1021/jp9046837.CrossRefPubMedGoogle Scholar
  54. 54.
    Bancelin S, Aimé C, Coradin T, Schanne-Klein M-C. In situ three-dimensional monitoring of collagen fibrillogenesis using SHG microscopy. Biomed Opt Express. 2012;3(6):1446–54. doi: 10.1364/BOE.3.001446.CrossRefPubMedPubMedCentralGoogle Scholar
  55. 55.
    Strupler M, Hernest M, Fligny C, Martin J-L, Tharaux P-L, Schanne-Klein M-C. Second harmonic microscopy to quantify renal interstitial fibrosis and arterial remodeling. J Biomed Opt. 2008;13(5):054041. doi: 10.1117/1.2981830.CrossRefPubMedGoogle Scholar
  56. 56.
    Sun W, Chang S, Tai DCS, et al. Nonlinear optical microscopy: use of second harmonic generation and two-photon microscopy for automated quantitative liver fibrosis studies. J Biomed Opt. 2008;13(6):064010. doi: 10.1117/1.3041159.CrossRefPubMedGoogle Scholar
  57. 57.
    Provenzano PP, Eliceiri KW, Campbell JM, Inman DR, White JG, Keely PJ. Collagen reorganization at the tumor-stromal interface facilitates local invasion. BMC Med. 2006;4(1):38. doi: 10.1186/1741-7015-4-38.CrossRefPubMedPubMedCentralGoogle Scholar
  58. 58.
    Nadiarnykh O, LaComb RB, Brewer MA, Campagnola PJ. Alterations of the extracellular matrix in ovarian cancer studied by Second Harmonic Generation imaging microscopy. BMC Cancer. 2010;10:94. doi: 10.1186/1471-2407-10-94.CrossRefPubMedPubMedCentralGoogle Scholar
  59. 59.
    Zhuo S, Chen J, Wu G, et al. Quantitatively linking collagen alteration and epithelial tumor progression by second harmonic generation microscopy. Appl Phys Lett. 2010;96(21):213704. doi: 10.1063/1.3441337.CrossRefGoogle Scholar
  60. 60.
    Latour G, Kowalczuk L, Savoldelli M, et al. Hyperglycemia-induced abnormalities in rat and human corneas: the potential of second harmonic generation microscopy. Georgakoudi I, ed. PLoS One. 2012;7(11):e48388. doi: 10.1371/journal.pone.0048388.CrossRefPubMedPubMedCentralGoogle Scholar
  61. 61.
    Tan H-Y, Sun Y, Lo W, et al. Multiphoton fluorescence and second harmonic generation microscopy for imaging infectious keratitis. J Biomed Opt. 2007;12(2):024013. doi: 10.1117/1.2717133.CrossRefPubMedGoogle Scholar
  62. 62.
    Matteini P, Cicchi R, Ratto F, et al. Thermal transitions of fibrillar collagen unveiled by second-harmonic generation microscopy of corneal stroma. Biophys J. 2012;103(6):1179–87. doi: 10.1016/j.bpj.2012.07.055.CrossRefPubMedPubMedCentralGoogle Scholar
  63. 63.
    Donnelly E, Williams RM, Downs SA, Dickinson ME, Baker SP, van der Meulen MCH. Quasistatic and dynamic nanomechanical properties of cancellous bone tissue relate to collagen content and organization. J Mater Res. 2011;21(08):2106–17. doi: 10.1557/jmr.2006.0259.CrossRefGoogle Scholar
  64. 64.
    Lacomb R, Nadiarnykh O, Campagnola PJ. Quantitative second harmonic generation imaging of the diseased state osteogenesis imperfecta: experiment and simulation. Biophys J. 2008;94(11):4504–14. doi: 10.1529/biophysj.107.114405.CrossRefPubMedPubMedCentralGoogle Scholar
  65. 65.
    Le TT, Langohr IM, Locker MJ, Sturek M, Cheng J-X. Label-free molecular imaging of atherosclerotic lesions using multimodal nonlinear optical microscopy. J Biomed Opt. 2007;12(5):054007. doi: 10.1117/1.2795437.CrossRefPubMedPubMedCentralGoogle Scholar
  66. 66.
    Zoumi A, Lu X, Kassab GS, Tromberg BJ. Imaging coronary artery microstructure using second-harmonic and two-photon fluorescence microscopy. Biophys J. 2004;87(4):2778–86. doi: 10.1529/biophysj.104.042887.CrossRefPubMedPubMedCentralGoogle Scholar
  67. 67.
    König K. Clinical multiphoton tomography. J Biophotonics. 2008;1(1):13–23. doi: 10.1002/jbio.200710022.CrossRefPubMedGoogle Scholar
  68. 68.
    Koehler MJ, Hahn S, Preller A, et al. Morphological skin ageing criteria by multiphoton laser scanning tomography: non-invasive in vivo scoring of the dermal fibre network. Exp Dermatol. 2008;17(6):519–23. doi: 10.1111/j.1600-0625.2007.00669.x.CrossRefPubMedGoogle Scholar
  69. 69.
    Wu S, Li H, Zhang X, Chen WR, Wang Y-X. Character of skin on photo-thermal response and its regeneration process using second-harmonic generation microscopy. Lasers Med Sci. 2014;29(1):141–6. doi: 10.1007/s10103-013-1296-3.CrossRefPubMedGoogle Scholar
  70. 70.
    Yasui T, Takahashi Y, Fukushima S, et al. Observation of dermal collagen fiber in wrinkled skin using polarization-resolved second-harmonic-generation microscopy. Opt Express. 2009;17(2):912–23. Available at: http://www.ncbi.nlm.nih.gov/pubmed/19158906. Accessed 9 Jul 2014.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Mathias Strupler
    • 1
  • Romain Deterre
    • 2
  • Nadir Goulamhoussen
    • 1
  • Fouzi Benboujja
    • 2
  • Christopher J. Hartnick
    • 3
  • Caroline Boudoux
    • 1
  1. 1.Department of Engineering PhysicsEcole Polytechnique MontréalMontrealCanada
  2. 2.Department of Biomedical EngineeringEcole PolytechniqueMontrealCanada
  3. 3.Department of Otolaryngology, Harvard Medical SchoolMassachusetts Eye and Ear InfirmaryBostonUSA

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