How many photons are needed for FRET imaging?

Biomedical Optics Express
Alessandro Esposito

Abstract

Förster resonance energy transfer (FRET) imaging is an essential analytical method in biomedical research. The limited photon-budget experimentally available, however, imposes compromises between spatiotemporal and biochemical resolutions, photodamage and phototoxicity. The study of photon-statistics in biochemical imaging is thus important in guiding the efficient design of instrumentation and assays. Here, we show a comparative analysis of photon-statistics in FRET imaging demonstrating how the precision of FRET imaging varies vastly with imaging parameters. Therefore, we provide analytical and numerical tools for assay optimization. Fluorescence lifetime imaging microscopy (FLIM) is a very robust technique with excellent photon-efficiencies. However, we show that also intensity-based FRET imaging can reach high precision by utilizing information from both donor and acceptor fluorophores.

References

Aug 1, 1967·Proceedings of the National Academy of Sciences of the United States of America·L Stryer, R P Haugland
Mar 1, 1996·Nature Biotechnology·S Tyagi, F R Kramer
Dec 24, 2002·Biophysical Journal·Adam HoppeJoel A Swanson
Jan 25, 2003·Methods : a Companion to Methods in Enzymology·Masilamani ElangovanAmmasi Periasamy
Feb 7, 2003·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·Johan Philip, Kjell Carlsson
May 29, 2003·Biophysical Journal·Claude Berney, Gaudenz Danuser
Jul 26, 2003·Journal of Biomedical Optics·Enrico GrattonNicholas Barry
Nov 5, 2003·Nature Biotechnology·Elizabeth A Jares-Erijman, Thomas M Jovin
Jun 11, 2004·Biophysical Journal·Lucas P Watkins, Haw Yang
Sep 8, 2004·Microscopy Research and Technique·Richard A Neher, Erwin Neher
Sep 24, 2004·International Review of Cytology·Gertrude Bunt, Fred S Wouters
Mar 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Sundar GanesanFred S Wouters
Mar 16, 2006·Microscopy Research and Technique·Horst WallrabeMargarida Barroso
Aug 23, 2006·Microscopy Research and Technique·Serge PeletPeter T C So
Sep 5, 2006·Current Opinion in Chemical Biology·Elizabeth A Jares-Erijman, Thomas M Jovin
Oct 4, 2007·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·Alessandro EspositoFred S Wouters
Oct 9, 2007·Biophysical Journal·Jakub WlodarczykErwin Neher
Nov 6, 2007·Biophysical Journal·Michelle A DigmanEnrico Gratton
Nov 17, 2007·Microscopy Research and Technique·Ryan A ColyerEnrico Gratton
Feb 5, 2008·Journal of the Optical Society of America. A, Optics, Image Science, and Vision·Alan ElderClemens F Kaminski
Apr 2, 2010·Chemical Reviews·Mikhail Y Berezin, Samuel Achilefu
Sep 22, 2011·Journal of Microscopy·F FereidouniH C Gerritsen
Dec 4, 2012·Biophysical Journal·André ZeugEvgeni G Ponimaskin
Nov 10, 2013·PloS One·Alessandro EspositoAshok R Venkitaraman
Apr 4, 2015·Optics Express·Nikola KrstajićRobert Henderson
Sep 15, 2015·Optics Express·Marina PopleteevaAlessandro Esposito
Sep 21, 2016·Sensors·Bryce T BajarJun Chu
Apr 25, 2017·Methods : a Companion to Methods in Enzymology·James R W ConwayPaul Timpson
Aug 23, 2018·Optics Express·Ivan Michel AntolovicEdoardo Charbon
Aug 20, 2019·Methods and Applications in Fluorescence·Andrew L TrinhAlessandro Esposito
Sep 13, 2019·Optics Express·Dorian BouchetIgnacio Izeddin

❮ Previous
Next ❯

Methods Mentioned

BETA
FRET
biosensors

Software Mentioned

FLIM
seFRET
TCSPC
dFRET
Mathematica
Matlab

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.

Related Papers

Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme
Hiroshi Oda
Science's STKE : Signal Transduction Knowledge Environment
Steven S VogelSrinagesh V Koushik
BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology
Sergi Padilla-Parra, Marc Tramier
© 2021 Meta ULC. All rights reserved