Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane

@article{Jinno2014EngineeringAG,
  title={Engineering a genetically-encoded SHG chromophore by electrostatic targeting to the membrane},
  author={Yuka Jinno and Keiko Shoda and Emiliano Rial-Verde and Rafael Yuste and Atsushi Miyawaki and Hidekazu Tsutsui},
  journal={Frontiers in Molecular Neuroscience},
  year={2014},
  volume={7}
}
Although second harmonic generation (SHG) microscopy provides unique imaging advantages for voltage imaging and other biological applications, genetically-encoded SHG chromophores remain relatively unexplored. SHG only arises from non-centrosymmetric media, so an anisotropic arrangement of chromophores is essential to provide strong SHG signals. Here, inspired by the mechanism by which K-Ras4B associates with plasma membranes, we sought to achieve asymmetric arrangements of chromophores at the… 

Figures from this paper

Application of Second Harmonic Imaging Microscopy in Biological Studies
The fact that some crystalline substances can generate the second harmonic (SH) – light of twice the frequency – under intense irradiation was discovered soon after lasers were available. Later, it
Non-invasive membrane potential measurements using endogenous markers
TLDR
The vibrational Stark spectroscopy is evaluated and it is demonstrated that it has sufficient sensitivity to assess physiologically significant variations of the membrane potential.
Harmonic Generation Microscopy 2.0: New Tricks Empowering Intravital Imaging for Neuroscience
TLDR
The fundamental contrast mechanisms are reviewed as they encode novel information including molecular origin, spectroscopy, functional probes, and image analysis, which lay foundations for promising future applications in neuroscience.
Optical second-harmonic images of sacran megamolecule aggregates.
We have detected a second-order nonlinear optical response from aggregates of the ampholytic megamolecular polysaccharide sacran extracted from cyanobacterial biomaterials by using optical
Genetic voltage indicators
TLDR
Although genetic voltage indicators could revolutionize neuroscience, there are still significant challenges, particularly two-photon performance, which may require cross-disciplinary collaborations, team effort, and sustained support by large-scale research initiatives.

References

SHOWING 1-10 OF 53 REFERENCES
A variant of yellow fluorescent protein with fast and efficient maturation for cell-biological applications
TLDR
The development of an improved version of YFP named Venus, which contains a novel mutation, F46L, which at 37°C greatly accelerates oxidation of the chromophore, the rate-limiting step of maturation and will enable fluorescent labelings that were not possible before.
Second-harmonic generation in GFP-like proteins.
TLDR
These properties are essential for the practical use of Dronpa or other GFP-like fluorescent proteins as second-order nonlinear fluorophores for symmetry-sensitive nonlinear microscopy imaging and as nonlinear optical sensors for electrophysiological processes.
Crystal Structure of Venus, a Yellow Fluorescent Protein with Improved Maturation and Reduced Environmental Sensitivity*
TLDR
The rearrangement of several side chains near the chromophore, initiated by the F46L mutation, was found to improve maturation at 37 °C by removing steric and energetic constraints, which may hinder folding of the polypeptide chain, and by accelerating the oxidation of the Cα–Cβ bond of Tyr66 during chromophores formation.
Video-rate nonlinear microscopy of neuronal membrane dynamics with genetically encoded probes.
TLDR
Comparison with second-harmonic generation by membranes stained with a synthetic styryl dye suggested that the genetically encoded chromophore arrangement lacked the orientational anisotropy and/or dipole density required for efficient coherent scattering of the incident optical field.
Anomalous negative fluorescence anisotropy in yellow fluorescent protein (YFP 10C): quantitative analysis of FRET in YFP dimers.
TLDR
It is proposed that the negative anisotropy of YFP 10C depends on protein concentration in the low micromolar range that was not expected, and is a result of unidirectional Förster resonance energy transfer in a dimer of Y FP, with the donor chromophore in the neutral form and the acceptor chromophores in the anionic form.
Mechanisms of membrane potential sensing with second-harmonic generation microscopy.
TLDR
Two mechanisms are found to contribute to the voltage response of a novel second-harmonic generation (SHG) marker: an electro-optic-induced alteration of the molecular hyperpolarizability and an electric-field-induced altered of the degree of molecular alignment.
Second-harmonic imaging microscopy of living cells.
TLDR
Second harmonic generation is complementary to TPEF in that it uses a different contrast mechanism and is most easily detected in the transmitted light optical path, and has the ability to image highly ordered structural proteins without any exogenous labels.
...
1
2
3
4
5
...