Repository logo
 

How many photons are needed for FRET imaging?

Accepted version
Peer-reviewed

Type

Article

Change log

Authors

Esposito, Alessandro  ORCID logo  https://orcid.org/0000-0002-5051-091X

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.

Description

Keywords

5102 Atomic, Molecular and Optical Physics, 51 Physical Sciences, Bioengineering

Journal Title

Biomed Opt Express

Conference Name

Journal ISSN

2156-7085
2156-7085

Volume Title

11

Publisher

Optica Publishing Group

Rights

All rights reserved
Sponsorship
MRC (unknown)
MRC (4050551988)
Medical Research Council (MC_UU_12022/1)
MRC (MC_UU_12022/8)