First-Principles Study of One- and Two-Photon Absorption of the H-Bonding Complexes from Monomeric Red Fluorescent Proteins with Large Stokes Shifts

2011 ◽  
Vol 115 (36) ◽  
pp. 10750-10757 ◽  
Author(s):  
Min-Yi Zhang ◽  
Jin-Yun Wang ◽  
Chen Sheng Lin ◽  
Wen-Dan Cheng
Keyword(s):  
First Principles ◽  
Fluorescent Proteins ◽  
Photon Absorption ◽  
Two Photon Absorption ◽  
Two Photon ◽  
First Principles Study ◽  
Red Fluorescent Proteins ◽  
Stokes Shifts

scholarly journals Multiphoton Bleaching of Red Fluorescent Proteins and the Ways to Reduce It

2022 ◽  
Vol 23 (2) ◽  
pp. 770
Author(s):  
Mikhail Drobizhev ◽  
Rosana S. Molina ◽  
Jacob Franklin
Keyword(s):  
Chemical Structure ◽  
Fluorescent Proteins ◽  
Multiphoton Ionization ◽  
Excitation Power ◽  
Photon Absorption ◽  
Green Fluorescent Proteins ◽  
Two Photon Absorption ◽  
Two Photon ◽  
Red Fluorescent Proteins ◽  
Green Fluorescent

Red fluorescent proteins and biosensors built upon them are potentially beneficial for two-photon laser microscopy (TPLM) because they can image deeper layers of tissue, compared to green fluorescent proteins. However, some publications report on their very fast photobleaching, especially upon excitation at 750–800 nm. Here we study the multiphoton bleaching properties of mCherry, mPlum, tdTomato, and jREX-GECO1, measuring power dependences of photobleaching rates K at different excitation wavelengths across the whole two-photon absorption spectrum. Although all these proteins contain the chromophore with the same chemical structure, the mechanisms of their multiphoton bleaching are different. The number of photons required to initiate a photochemical reaction varies, depending on wavelength and power, from 2 (all four proteins) to 3 (jREX-GECO1) to 4 (mCherry, mPlum, tdTomato), and even up to 8 (tdTomato). We found that at sufficiently low excitation power P, the rate K often follows a quadratic power dependence, that turns into higher order dependence (K~Pα with α > 2) when the power surpasses a particular threshold P*. An optimum intensity for TPLM is close to the P*, because it provides the highest signal-to-background ratio and any further reduction of laser intensity would not improve the fluorescence/bleaching rate ratio. Additionally, one should avoid using wavelengths shorter than a particular threshold to avoid fast bleaching due to multiphoton ionization.

scholarly journals Absolute Two-Photon Absorption Spectra and Two-Photon Brightness of Orange and Red Fluorescent Proteins

2009 ◽  
Vol 113 (4) ◽  
pp. 855-859 ◽  
Author(s):  
M. Drobizhev ◽  
S. Tillo ◽  
N. S. Makarov ◽  
T. E. Hughes ◽  
A. Rebane
Keyword(s):  
Absorption Spectra ◽  
Fluorescent Proteins ◽  
Photon Absorption ◽  
Two Photon Absorption ◽  
Two Photon ◽  
Red Fluorescent Proteins

scholarly journals Absolute Two-photon Absorption Spectra Of Orange And Red Fluorescent Proteins

2009 ◽  
Vol 96 (3) ◽  
pp. 400a-401a
Author(s):  
Mikhail Drobizhev ◽  
Shane Tillo ◽  
Nikolay Makarov ◽  
Aleksander Rebane ◽  
Thomas E. Hughes
Keyword(s):  
Absorption Spectra ◽  
Fluorescent Proteins ◽  
Photon Absorption ◽  
Two Photon Absorption ◽  
Two Photon ◽  
Red Fluorescent Proteins

scholarly journals Two-Photon Absorption Properties of Chromophores of a Few Fluorescent Proteins: a Theoretical Investigation

2016 ◽  
Vol 32 (1) ◽  
pp. 301-312
Author(s):  
Chuan-Xiang YE ◽  
◽  
Hui-Li MA ◽  
Wan-Zhen LIANG ◽  
◽  
...  
Keyword(s):  
Theoretical Investigation ◽  
Fluorescent Proteins ◽  
Photon Absorption ◽  
Absorption Properties ◽  
Two Photon Absorption ◽  
Two Photon

Resonance Enhancement of Two-Photon Absorption in Fluorescent Proteins

2007 ◽  
Vol 111 (50) ◽  
pp. 14051-14054 ◽  
Author(s):  
M. Drobizhev ◽  
N. S. Makarov ◽  
T. Hughes ◽  
A. Rebane
Keyword(s):  
Fluorescent Proteins ◽  
Photon Absorption ◽  
Two Photon Absorption ◽  
Two Photon ◽  
Resonance Enhancement
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