TY - JOUR
T1 - A FLIM Microscopy Based on Acceptor-Detected Förster Resonance Energy Transfer
AU - Delgadillo, Roberto F.
AU - Carnes, Katie A.
AU - Zaleta-Rivera, Kathia
AU - Olmos, Omar
AU - Parkhurst, Lawrence J.
N1 - Funding Information:
The work was supported by National Institutes of Health Grants GM59346 and RR015468 to LJP; CONACYT-Mexico postdoctoral, SNI fellowships (130994,162809, SNI75487), and the Government of Veracruz-Mexico fellowships to R.F.D.
Publisher Copyright:
©
PY - 2021
Y1 - 2021
N2 - Time-resolved donor-detected Förster resonance energy transfer (trDDFRET) allows the observation of molecular interactions of dye-labeled biomolecules in the â10-100 Å region. However, we can observe longer-range interactions when using time-resolved acceptor-detected FRET (trADFRET), since the signal/noise ratio can be improved when observing the acceptor emission. Therefore, we propose a new methodology based on trADFRET to construct a new fluorescence lifetime microscopy (FLIM-trADFRET) technique to observe biological machinery in the range of 100-300 Å in vivo, the last frontier in biomolecular medicine. The integrated trADFRET signal is extracted in such a way that noise is canceled, and more photons are collected, even though trADFRET and trDDFRET have the same rate of transfer. To assess our new methodology, proof of concept was demonstrated with a set of well-defined DNA scaffolds.
AB - Time-resolved donor-detected Förster resonance energy transfer (trDDFRET) allows the observation of molecular interactions of dye-labeled biomolecules in the â10-100 Å region. However, we can observe longer-range interactions when using time-resolved acceptor-detected FRET (trADFRET), since the signal/noise ratio can be improved when observing the acceptor emission. Therefore, we propose a new methodology based on trADFRET to construct a new fluorescence lifetime microscopy (FLIM-trADFRET) technique to observe biological machinery in the range of 100-300 Å in vivo, the last frontier in biomolecular medicine. The integrated trADFRET signal is extracted in such a way that noise is canceled, and more photons are collected, even though trADFRET and trDDFRET have the same rate of transfer. To assess our new methodology, proof of concept was demonstrated with a set of well-defined DNA scaffolds.
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U2 - 10.1021/acs.analchem.0c04492
DO - 10.1021/acs.analchem.0c04492
M3 - Article
C2 - 33691398
AN - SCOPUS:85103466963
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
ER -