scholarly journals Describing Two-Photon Absorptivity of Fluorescent Proteins with a New Vibronic Coupling Mechanism

2012 ◽  
Vol 116 (5) ◽  
pp. 1736-1744 ◽  
Author(s):  
M. Drobizhev ◽  
N. S. Makarov ◽  
S. E. Tillo ◽  
T. E. Hughes ◽  
A. Rebane
Keyword(s):  
Fluorescent Proteins ◽  
Vibronic Coupling ◽  
Coupling Mechanism ◽  
Two Photon

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 Directionality of Two-Photon Excitation in Representative Fluorescent Proteins

2018 ◽  
Vol 114 (3) ◽  
pp. 171a-172a
Author(s):  
Olga Rybakova ◽  
Stepan Timr ◽  
Josef Lazar
Keyword(s):  
Fluorescent Proteins ◽  
Two Photon ◽  
Photon Excitation ◽  
Two Photon Excitation

scholarly journals Two-photon directed evolution of green fluorescent proteins

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Caleb R. Stoltzfus ◽  
Lauren M. Barnett ◽  
Mikhail Drobizhev ◽  
Geoffrey Wicks ◽  
Alexander Mikhaylov ◽  
...  
Keyword(s):  
Directed Evolution ◽  
Fluorescent Proteins ◽  
Green Fluorescent Proteins ◽  
Two Photon ◽  
Green Fluorescent

scholarly journals Development of an Immunologically Tolerated Combination of Fluorescent Proteins for In vivo Two-photon Imaging

2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Selamawit Gossa ◽  
Debasis Nayak ◽  
Bernd H. Zinselmeyer ◽  
Dorian B. McGavern
Keyword(s):  
Fluorescent Proteins ◽  
Two Photon ◽  
Photon Imaging ◽  
Two Photon Imaging

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

Two-photon dual-color imaging using fluorescent proteins

2008 ◽  
Vol 5 (5) ◽  
pp. 373-374 ◽  
Author(s):  
Hiroyuki Kawano ◽  
Takako Kogure ◽  
Yukiko Abe ◽  
Hideaki Mizuno ◽  
Atsushi Miyawaki
Keyword(s):  
Fluorescent Proteins ◽  
Color Imaging ◽  
Two Photon ◽  
Dual Color
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