Analytical and Bioanalytical Chemistry

, Volume 411, Issue 19, pp 4861–4871 | Cite as

High-resolution and high-repetition-rate vibrational sum-frequency generation spectroscopy of one- and two-component phosphatidylcholine monolayers

  • Freeda Yesudas
  • Mark Mero
  • Janina Kneipp
  • Zsuzsanna HeinerEmail author
Research Paper
Part of the following topical collections:
  1. Young Investigators in (Bio-)Analytical Chemistry


We present broadband vibrational sum-frequency generation (VSFG) spectra of Langmuir-Blodgett monolayers of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and different mixtures of them as model systems of pulmonary surfactants. The systematic study explored the dependence of the vibrational spectra as a function of surface tension and mixture ratio in various polarization combinations. The extremely short acquisition time and the high spectral resolution of our recently developed spectrometer helped minimize sample degradation under ambient conditions throughout the duration of the measurement and allowed the detection of previously unseen vibrational bands with unprecedented signal-to-noise ratio. The dramatically improved capability to record reliable vibrational spectra together with the label-free nature of the VSFG method provides direct access to native lipid structure and dynamics directly in the monolayer. The resulting data deliver quantitative information for structural analysis of multi-component phospholipid monolayers and may aid in the development of new synthetic pulmonary surfactants.


Lung surfactants 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) Broadband vibrational sum-frequency generation spectroscopy Two-component phosphatidylcholine Langmuir-Blodgett monolayer 



We thank K. Balasubramanian (SALSA, Humboldt-Universität zu Berlin) and R. M. Iost for providing support in the sample cleaning processes.

Funding information

This project is financed by the Deutsche Forschungsgemeinschaft (DFG) project GSC 1013 SALSA. F.Y. is grateful for the support by a fellowship in SALSA. Z. H. acknowledges the funding of her Julia Lermontova fellowship by GSC 1013 SALSA.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

216_2019_1690_MOESM1_ESM.pdf (466 kb)
ESM 1 (PDF 466 kb)


  1. 1.
    Goerke J. Pulmonary surfactant: functions and molecular composition. Biochim Biophys Acta (BBA) - Mol Basis Dis. 1998;1408(2):79–89. Scholar
  2. 2.
    Wüstneck R, Perez-Gil J, Wüstneck N, Cruz A, Fainerman VB, Pison U. Interfacial properties of pulmonary surfactant layers. Adv Colloid Interf Sci. 2005;117(1):33–58. Scholar
  3. 3.
    Yu S, Harding PGR, Smith N, Possmayer F. Bovine pulmonary surfactant: chemical composition and physical properties. Lipids. 1983;18(8):522–9. Scholar
  4. 4.
    Ramanathan R. Animal-derived surfactants: where are we? The evidence from randomized, controlled clinical trials. J Perinatol. 2009;29:S38. Scholar
  5. 5.
    Sardesai S, Biniwale M, Wertheimer F, Garingo A, Ramanathan R. Evolution of surfactant therapy for respiratory distress syndrome: past, present, and future. Pediatr Res. 2016;81:240. Scholar
  6. 6.
    Pfister RH, Soll R, Wiswell TE. Cochrane review: Protein containing synthetic surfactant versus animal derived surfactant extract for the prevention and treatment of respiratory distress syndrome. Evid Based Child Health. 2010;5(1):17–51. Scholar
  7. 7.
    Halliday HL. Surfactants: past, present and future. J Perinatol. 2008;28:S47. Scholar
  8. 8.
    Olżyńska A, Zubek M, Roeselova M, Korchowiec J, Cwiklik L. Mixed DPPC/POPC monolayers: all-atom molecular dynamics simulations and Langmuir monolayer experiments. Biochim Biophys Acta Biomembr. 2016;1858(12):3120–30. Scholar
  9. 9.
    Wang H-F, Velarde L, Gan W, Fu L. Quantitative sum-frequency generation vibrational spectroscopy of molecular surfaces and interfaces: lineshape, polarization, and orientation. Annu Rev Phys Chem. 2015;66(1):189–216. Scholar
  10. 10.
    Wang H-F, Gan W, Lu R, Rao Y, Wu B-H. Quantitative spectral and orientational analysis in surface sum frequency generation vibrational spectroscopy (SFG-VS). Int Rev Phys Chem. 2005;24(2):191–256. Scholar
  11. 11.
    Baumler SM, Allen HC. Chapter 5. Vibrational spectroscopy of gas–liquid interfaces. In: Faust JA, House JE, editors. Physical chemistry of gas-liquid interfaces. Elsevier; 2018. p. 105–33.Google Scholar
  12. 12.
    Liu W, Wang Z, Fu L, Leblanc RM, Yan ECY. Lipid compositions modulate fluidity and stability of bilayers: characterization by surface pressure and sum frequency generation spectroscopy. Langmuir. 2013;29(48):15022–31. Scholar
  13. 13.
    Li B, Lu X, Han X, Wu F-G, Myers JN, Chen Z. Interfacial Fresnel coefficients and molecular structures of model cell membranes: from a lipid monolayer to a lipid bilayer. J Phys Chem C. 2014;118(49):28631–9. Scholar
  14. 14.
    Ma G, Allen HC. DPPC Langmuir monolayer at the air–water interface: probing the tail and head groups by vibrational sum frequency generation spectroscopy. Langmuir. 2006;22(12):5341–9. Scholar
  15. 15.
    Sung W, Seok S, Kim D, Tian CS, Shen YR. Sum-frequency spectroscopic study of Langmuir monolayers of lipids having oppositely charged headgroups. Langmuir. 2010;26(23):18266–72. Scholar
  16. 16.
    Nojima Y, Suzuki Y, Yamaguchi S. Weakly hydrogen-bonded water inside charged lipid monolayer observed with heterodyne-detected vibrational sum frequency generation spectroscopy. J Phys Chem C. 2017;121(4):2173–80. Scholar
  17. 17.
    Liljeblad JFD, Bulone V, Rutland MW, Johnson CM. Supported phospholipid monolayers. The molecular structure investigated by vibrational sum frequency spectroscopy. J Phys Chem C. 2011;115(21):10617–29. Scholar
  18. 18.
    Li B, Li X, Ma Y-H, Han X, Wu F-G, Guo Z, et al. Sum frequency generation of interfacial lipid monolayers shows polarization dependence on experimental geometries. Langmuir. 2016;32(28):7086–95. Scholar
  19. 19.
    Feng R-J, Li X, Zhang Z, Lu Z, Guo Y. Spectral assignment and orientational analysis in a vibrational sum frequency generation study of DPPC monolayers at the air/water interface. J Chem Phys. 2016;145(24):244707. Scholar
  20. 20.
    Smits M, Sovago M, Wurpel GWH, Kim D, Müller M, Bonn M. Polarization-resolved broad-bandwidth sum-frequency generation spectroscopy of monolayer relaxation. J Phys Chem C. 2007;111(25):8878–83. Scholar
  21. 21.
    Liljeblad JFD, Bulone V, Tyrode E, Rutland MW, Johnson CM. Phospholipid monolayers probed by vibrational sum frequency spectroscopy: instability of unsaturated phospholipids. Biophys J. 2010;98(10):L50–L2. Scholar
  22. 22.
    Velarde L, Wang H-F. Unified treatment and measurement of the spectral resolution and temporal effects in frequency-resolved sum-frequency generation vibrational spectroscopy (SFG-VS). Phys Chem Chem Phys. 2013;15(46):19970–84. Scholar
  23. 23.
    Li Y, Feng R, Lin L, Liu M, Guo Y, Zhang Z. Ordering effects of cholesterol on sphingomyelin monolayers investigated by high-resolution broadband sum-frequency generation vibrational spectroscopy. Chin Chem Lett. 2018;29(3):357–60. Scholar
  24. 24.
    Heiner Z, Petrov V, Mero M. Compact, high-repetition-rate source for broadband sum-frequency generation spectroscopy. APL Photonics. 2017;2(6):066102. Scholar
  25. 25.
    Yesudas F, Mero M, Kneipp J, Heiner Z. Vibrational sum-frequency generation spectroscopy of lipid bilayers at repetition rates up to 100 kHz. J Chem Phys. 2018;148(10):104702. Scholar
  26. 26.
    Roberts G. Langmuir-Blodgett films. Boston: Springer; 1990.CrossRefGoogle Scholar
  27. 27.
    Motschmann H, Mohwald H. Langmuir–Blodgett films. In: Holmberg K, editor. Handbook of applied surface and colloid chemistry. Wiley; 2001.Google Scholar
  28. 28.
    Tredgold RH. The physics of Langmuir-Blodgett films. Rep Prog Phys. 1987;50(12):1609.CrossRefGoogle Scholar
  29. 29.
    Raoult F, Boscheron ACL, Husson D, Sauteret C, Modena A, Malka V, et al. Efficient generation of narrow-bandwidth picosecond pulses by frequency doubling of femtosecond chirped pulses. Opt Lett. 1998;23(14):1117–9. Scholar
  30. 30.
    Nejbauer M, Radzewicz C. Efficient spectral shift and compression of femtosecond pulses by parametric amplification of chirped light. Opt Express. 2012;20(3):2136–42. Scholar
  31. 31.
    Qiao L, Ge A, Liang Y, Ye S. Oxidative degradation of the monolayer of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) in low-level ozone. J Phys Chem B. 2015;119(44):14188–99. Scholar
  32. 32.
    Liu J, Conboy JC. Structure of a gel phase lipid bilayer prepared by the Langmuir–Blodgett/Langmuir-Schaefer method characterized by sum-frequency vibrational spectroscopy. Langmuir. 2005;21(20):9091–7. Scholar
  33. 33.
    Lu R, Gan W, Wu B-h, Chen H, Wang H-f. Vibrational polarization spectroscopy of CH stretching modes of the methylene group at the vapor/liquid interfaces with sum frequency generation. J Phys Chem B. 2004;108(22):7297–306. Scholar
  34. 34.
    Lu R, Gan W, Wu B-h, Zhang Z, Guo Y, Wang H-f. C–H stretching vibrations of methyl, methylene and methine groups at the vapor/alcohol (n = 1–8) interfaces. J Phys Chem B. 2005;109(29):14118–29. Scholar
  35. 35.
    Velarde L, Wang H-f. Capturing inhomogeneous broadening of the –CN stretch vibration in a Langmuir monolayer with high-resolution spectra and ultrafast vibrational dynamics in sum-frequency generation vibrational spectroscopy (SFG-VS). J Chem Phys. 2013;139(8):084204. Scholar
  36. 36.
    Velarde L, Zhang X-y, Lu Z, Joly AG, Wang Z, Wang H-f. Communication: Spectroscopic phase and lineshapes in high-resolution broadband sum frequency vibrational spectroscopy: resolving interfacial inhomogeneities of “identical” molecular groups. J Chem Phys. 2011;135(24):241102. Scholar
  37. 37.
    Feng R-j, Lin L, Li Y-y, Liu M-h, Guo Y, Zhang Z. Effect of Ca2+ to sphingomyelin investigated by sum frequency generation vibrational spectroscopy. Biophys J. 2017;112(10):2173–83. Scholar
  38. 38.
    Fu L, Chen S-L, Wang H-F. Validation of spectra and phase in sub-1 cm−1 resolution sum-frequency generation vibrational spectroscopy through internal heterodyne phase-resolved measurement. J Phys Chem B. 2016;120(8):1579–89. Scholar
  39. 39.
    Mifflin AL, Velarde L, Ho J, Psciuk BT, Negre CFA, Ebben CJ, et al. Accurate line shapes from sub-1 cm−1 resolution sum frequency generation vibrational spectroscopy of α-pinene at room temperature. J Phys Chem A. 2015;119(8):1292–302. Scholar
  40. 40.
    MacPhail RA, Strauss HL, Snyder RG, Elliger CA. Carbon-hydrogen stretching modes and the structure of n-alkyl chains. 2. Long, all-trans chains. J Phys Chem. 1984;88(3):334–41. Scholar
  41. 41.
    Ward RN, Duffy DC, Davies PB, Bain CD. Sum-frequency spectroscopy of surfactants adsorbed at a flat hydrophobic surface. J Phys Chem. 1994;98(34):8536–42. Scholar
  42. 42.
    Conboy JC, Messmer MC, Richmond GL. Investigation of surfactant conformation and order at the liquid–liquid interface by total internal reflection sum-frequency vibrational spectroscopy. J Phys Chem. 1996;100(18):7617–22. Scholar
  43. 43.
    Miranda PB, Pflumio V, Saijo H, Shen YR. Chain–chain interaction between surfactant monolayers and alkanes or alcohols at solid/liquid interfaces. J Am Chem Soc. 1998;120(46):12092–9. Scholar
  44. 44.
    Watry MR, Tarbuck TL, Richmond GL. Vibrational sum-frequency studies of a series of phospholipid monolayers and the associated water structure at the vapor/water interface. J Phys Chem B. 2003;107(2):512–8. Scholar
  45. 45.
    Backus EHG, Bonn D, Cantin S, Roke S, Bonn M. Laser-heating-induced displacement of surfactants on the water surface. J Phys Chem B. 2012;116(9):2703–12. Scholar
  46. 46.
    Lambert AG, Davies PB, Neivandt DJ. Implementing the theory of sum frequency generation vibrational spectroscopy: a tutorial review. Appl Spectrosc Rev. 2005;40(2):103–45. Scholar
  47. 47.
    Kim J, Kim G, Cremer PS. Investigations of water structure at the solid/liquid interface in the presence of supported lipid bilayers by vibrational sum frequency spectroscopy. Langmuir. 2001;17(23):7255–60. Scholar
  48. 48.
    Chen X, Hua W, Huang Z, Allen HC. Interfacial water structure associated with phospholipid membranes studied by phase-sensitive vibrational sum frequency generation spectroscopy. J Am Chem Soc. 2010;132(32):11336–42. Scholar
  49. 49.
    Mondal JA, Nihonyanagi S, Yamaguchi S, Tahara T. Structure and orientation of water at charged lipid monolayer/water interfaces probed by heterodyne-detected vibrational sum frequency generation spectroscopy. J Am Chem Soc. 2010;132(31):10656–7. Scholar
  50. 50.
    Doǧangün M, Ohno PE, Liang D, McGeachy AC, Bé AG, Dalchand N, et al. Hydrogen-bond networks near supported lipid bilayers from vibrational sum frequency generation experiments and atomistic simulations. J Phys Chem B. 2018;122(18):4870–9. Scholar
  51. 51.
    Mondal JA, Nihonyanagi S, Yamaguchi S, Tahara T. Three distinct water structures at a zwitterionic lipid/water interface revealed by heterodyne-detected vibrational sum frequency generation. J Am Chem Soc. 2012;134(18):7842–50. Scholar
  52. 52.
    Notter RH, Tabak SA, Holcomb S, Mavis RD. Postcollapse dynamic surface pressure relaxation in binary surface films containing dipalmitoyl phosphatidylcholine. J Colloid Interface Sci. 1980;74(2):370–7. Scholar
  53. 53.
    Ma G, Liu J, Fu L, Yan ECY. Probing water and biomolecules at the air–water interface with a broad bandwidth vibrational sum frequency generation spectrometer from 3800 to 900 cm−1. Appl Spectrosc. 2009;63(5):528–37.CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Freeda Yesudas
    • 1
    • 2
  • Mark Mero
    • 3
  • Janina Kneipp
    • 1
    • 2
  • Zsuzsanna Heiner
    • 1
    • 2
    Email author
  1. 1.School of Analytical Sciences AdlershofHumboldt Universität zu BerlinBerlinGermany
  2. 2.Department of ChemistryHumboldt Universität zu BerlinBerlinGermany
  3. 3.Max Born Institute for Nonlinear Optics and Short Pulse SpectroscopyBerlinGermany

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