Two-photon excitation action cross-sections of the autofluorescent proteins

Blab, Gerhard A.; Lommerse, Piet H.M.; Cognet, Laurent; Harms, Gregory S.; Schmidt, Thomas

doi:10.1016/s0009-2614(01)01282-9

Cited by 120 publications

(137 citation statements)

References 24 publications

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“…In the case of two-photon excitation η/η flavine was close to unity. As a result, One find R = 36 at 860 nm for eCFP, R = 467 at 920 nm for eGFP and R = 566 at 960 nm for eYFP [46]. As predicted, the autofluorescence should not be a limitation to the observation of single autofluorescent proteins in living cells using two photon excitation.…”

Section: Two-photon Excitationsupporting

confidence: 56%

“…This particular applicability for depth discrimination in thick tissues results from the fact that fluorescence signal intensity is highly dependent on the degree of photon flux, which decreases rapidly as a function of distance from the focal plane. Measurements of the one and two-photon absorption cross-section of XFPs in bulk [46] show a super-linear scaling behavior which has been previously described only for small organic molecules. Furthermore, the two-photon absorption spectra closely follow those measured for one-photon absorption.…”

Section: Two-photon Excitationmentioning

confidence: 68%

“…However, a significant blue shift is found in the two-photon absorption cross-section of the autofluorescent proteins which can be attributed to the participation of a vibrational mode in the two-photon absorption process. In a similar way to the one photon case, one can generalize the excitation-wavelength dependent relative detection yield for the autofluorescence proteins to the two photon case [46] where the experimental detection efficiencies for the various fluorophores, η, and the absorption cross-sections, σ OPE and σ TPE , for the one-or two-photon case are considered. In the case of two-photon excitation η/η flavine was close to unity.…”

Section: Two-photon Excitationmentioning

confidence: 99%

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Fluorescence microscopy of single autofluorescent proteins for cellular biology

Cognet¹,

Coussen

Choquet

et al. 2002

Comptes Rendus. Physique

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In this paper we review the applicability of autofluorescent proteins for singlemolecule imaging in biology. The photophysical characteristics of several mutants of the Green Fluorescent Protein (GFP) and those of DsRed are compared and critically discussed for their use in cellular biology. The alternative use of twophoton excitation at the single-molecule level or Fluorescence Correlation Spectroscopy is envisaged for the study of individual autofluorescent proteins. Single-molecule experiments performed in live cells using eGFP and preferably eYFP fusion proteins are reviewed. Finally, the first use at the single-molecule level of citrine, a more photostable variant of the eYFP is reported when fused to a receptor for neurotransmitter in live cells.

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Section: Two-photon Excitationsupporting

confidence: 56%

Section: Two-photon Excitationmentioning

confidence: 68%

Section: Two-photon Excitationmentioning

confidence: 99%

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Fluorescence microscopy of single autofluorescent proteins for cellular biology

Cognet¹,

Coussen

Choquet

et al. 2002

Comptes Rendus. Physique

Self Cite

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“…Indeed, Spiess et al 13 do not report wavelengths below 435 nm (λ laser = 870 nm) and Blab et al 12 not below 375 nm (λ laser = 750 nm) for the TPA spectrum of YFP. Drobizhev et al report the TPA spectrum of Citrine (an optimized YFP variant) down to 600 nm.…”

Section: Discussionmentioning

confidence: 99%

Intermolecular charge transfer enhances two-photon absorption in yellow fluorescent protein

Beerepoot

Friese

Ruud

2014

Phys. Chem. Chem. Phys.

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We present a quantum chemical study of the two-photon absorption (TPA) properties of yellow fluorescent protein (YFP), a mutant of the extensively studied green fluorescent protein. The aromatic chromophore of YFP has a π-stacking interaction with the aromatic ring of a tyrosine residue (Tyr203) in a parellel-displaced structure with a distance of about 3.4Å. We study the TPA spectrum of the π-stacking system of YFP using the well-established Coulomb-attenuated B3LYP density functional (CAM-B3LYP) and the second-order approximate coupled-cluster model CC2. This work presents both the first comprehensive study of the two-photon absorption spectrum of YFP and the largest-scale coupled-cluster calculation of two-photon absorption that has ever been performed. We analyze the intermolecular charge-transfer (ICT) transitions in this stacked system and show that the ICT transitions are an important mechanism for enhancing the TPA cross sections in YFP. We investigate the distance dependence of the ICT transitions and show that their TPA cross sections are strongly dependent on the separation of the aromatic moieties. This provides a means for tuning the TPA properties of YFP and other structurally related fluorescent proteins through molecular engineering.

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“…2, since the linear and two-photon absorption bands lie between 430-550 nm and 850-1030 nm, respectively [41]. It is of essential impor- tance to our experiment that there exists a significant shift between twice the one-photon and the two-photon excitation band maxima of EYFP, so that direct twophoton excitation of EYFP by irradiation at 1064 nm is avoided.…”

Section: Experiments a Experimental Methodsmentioning

confidence: 84%

Fluorescence excitation by enhanced plasmon upconversion under continuous wave illumination

Tașgın

Salakhutdinov

Kendziora

et al. 2016

Photonics and Nanostructures - Fundamentals and Applications

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We demonstrate effective background-free continuous wave nonlinear optical excitation of molecules that are sandwiched between asymmetrically constructed plasmonic gold nanoparticle clusters. We observe that near infrared photons are converted to visible photons through efficient plasmonic second harmonic generation. Our theoretical model and simulations demonstrate that Fano resonances may be responsible for being able to observe nonlinear conversion using a continuous wave light source. We show that nonlinearity enhancement of plasmonic nanostructures via coupled quantum mechanical oscillators such as molecules can be several orders larger as compared to their classical counterparts.

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Two-photon excitation action cross-sections of the autofluorescent proteins

Cited by 120 publications

References 24 publications

Fluorescence microscopy of single autofluorescent proteins for cellular biology

Fluorescence microscopy of single autofluorescent proteins for cellular biology

Intermolecular charge transfer enhances two-photon absorption in yellow fluorescent protein

Fluorescence excitation by enhanced plasmon upconversion under continuous wave illumination

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