NONLINEAR OPTICAL PROPERTIES OF mSTRAWBERRY AND mCHERRY FOR SECOND HARMONIC IMAGING

The second-order nonlinear optical properties of two monomeric red fluorescent proteins, mStrawberry and mCherry, have been experimentally determined by frequency-resolved femtosecond hyper-Rayleigh scattering. These proteins were found to exhibit a stronger nonlinear response than the previously described eGFP, eYFP and DsRed,1 confirming the trend that fluorophores with a more extended conjugated system, or a lower-energy band gap between ground and excited state, exhibit a larger static hyperpolarizability (β0). Furthermore, these experimental data were complemented with quantum chemical calculations. A discrepancy was observed between experimental and theoretical results, but this could be explained by the chromophore model extracted from the available X-ray diffraction data. While eGFP showed a larger dynamic experimental response (βHRS) due to the highest resonance enhancement, we measured an even higher signal for mCherry. Furthermore, mCherry also shows a better separation of the second harmonic signal and two-photon excited fluorescent signal, making this the preferred fluorescent protein for second harmonic imaging at 800 nm so far.

[1]  B. Champagne,et al.  Time-dependent density functional theory simulation of UV/visible absorption spectra of zirconocene catalysts , 2002 .

[2]  R. Tsien,et al.  green fluorescent protein , 2020, Catalysis from A to Z.

[3]  Dmitrij Rappoport,et al.  Photoinduced intramolecular charge transfer in 4-(dimethyl)aminobenzonitrile--a theoretical perspective. , 2004, Journal of the American Chemical Society.

[4]  J. Oudar,et al.  Hyperpolarizabilities of the nitroanilines and their relations to the excited state dipole moment , 1977 .

[5]  B. Champagne,et al.  Investigation of the UV/visible absorption spectra of merocyanine dyes using time-dependent density functional theory. , 2006, The journal of physical chemistry. A.

[6]  M. Drobizhev,et al.  Absolute two-photon absorption spectra and two-photon brightness of orange and red fluorescent proteins. , 2009, The journal of physical chemistry. B.

[7]  Koen Clays,et al.  Second-order nonlinear optical properties of fluorescent proteins for second-harmonic imaging , 2009 .

[8]  J. Tomasi,et al.  Quantum mechanical continuum solvation models. , 2005, Chemical reviews.

[9]  O. Shimomura,et al.  Structure of the chromophore of Aequorea green fluorescent protein , 1979 .

[10]  S. Karna,et al.  Frequency dependent nonlinear optical properties of molecules: Formulation and implementation in the HONDO program , 1991 .

[11]  Yuriko Aoki,et al.  Assessment of time-dependent density functional schemes for computing the oscillator strengths of benzene, phenol, aniline, and fluorobenzene. , 2007, The Journal of chemical physics.

[12]  Atsushi Miyawaki,et al.  Second-harmonic generation in GFP-like proteins. , 2008, Journal of the American Chemical Society.

[13]  Leslie M. Loew,et al.  Second-harmonic generation of biological interfaces: probing the membrane protein bacteriorhodopsin and imaging membrane potential around GFP molecules at specific sites in neuronal cells of C. elegans , 1999 .

[14]  S. Tretiak,et al.  Prediction of Two-Photon Absorption Properties for Organic Chromophores Using Time-Dependent Density-Functional Theory , 2004 .

[15]  Koen Clays,et al.  Hyper-Rayleigh scattering in the Fourier domain for higher precision: Correcting for multiphoton fluorescence with demodulation and phase data , 2001 .

[16]  R Y Tsien,et al.  Biochemistry, mutagenesis, and oligomerization of DsRed, a red fluorescent protein from coral. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Nathan C Shaner,et al.  Novel chromophores and buried charges control color in mFruits. , 2006, Biochemistry.

[18]  R Y Tsien,et al.  Wavelength mutations and posttranslational autoxidation of green fluorescent protein. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Rafael Yuste,et al.  A two-photon and second-harmonic microscope. , 2003, Methods.

[20]  J. Tomasi,et al.  The effects of solvation in the theoretical spectra of cationic dyes , 2005 .

[21]  O. Shimomura,et al.  Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. , 1962, Journal of cellular and comparative physiology.

[22]  Jacopo Tomasi,et al.  Molecular Interactions in Solution: An Overview of Methods Based on Continuous Distributions of the Solvent , 1994 .

[23]  K. Ruud,et al.  TDDFT diagnostic testing and functional assessment for triazene chromophores. , 2009, Physical chemistry chemical physics : PCCP.

[24]  Koen Clays,et al.  High-frequency demodulation of multi-photon fluorescence in hyper-Rayleigh scattering , 1998 .

[25]  R. Tsien,et al.  Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein , 2004, Nature Biotechnology.

[26]  R. Bartlett,et al.  Nuclear spin–spin coupling constants evaluated using many body methods , 1986 .

[27]  Walter Thiel,et al.  Benchmarks for electronically excited states: CASPT2, CC2, CCSD, and CC3. , 2008, The Journal of chemical physics.

[28]  M. Davidson,et al.  Advances in fluorescent protein technology , 2011, Journal of Cell Science.

[29]  Hideo Sekino,et al.  Frequency dependent nonlinear optical properties of molecules , 1986 .

[30]  J Michiels,et al.  Identification of different emitting species in the red fluorescent protein DsRed by means of ensemble and single-molecule spectroscopy , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[31]  K. Hirao,et al.  A long-range-corrected time-dependent density functional theory. , 2004, The Journal of chemical physics.