High speed 2-photon fluorescence lifetime imaging of protein-protein interactions with a swept array microscope (Conference Presentation)

2020 ◽  
Author(s):  
Simon M. Ameer-Beg ◽  
Simon P. Poland ◽  
Thomas Kavanagh ◽  
James A. Levitt ◽  
Robert K. Henderson ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Protein Interactions ◽  
High Speed ◽  
Fluorescence Lifetime Imaging ◽  
Protein Protein Interactions ◽  
Lifetime Imaging ◽  
Photon Fluorescence

scholarly journals High-speed imaging of transient metabolic dynamics using two-photon fluorescence lifetime imaging microscopy

Optica ◽  
2018 ◽  
Vol 5 (10) ◽  
pp. 1290 ◽  
Author(s):  
Andrew J. Bower ◽  
Joanne Li ◽  
Eric J. Chaney ◽  
Marina Marjanovic ◽  
Darold R. Spillman ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
High Speed Imaging ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

scholarly journals Multiphoton time-domain fluorescence lifetime imaging microscopy: practical application to protein–protein interactions using global analysis

2008 ◽  
Vol 6 (suppl_1) ◽  
Author(s):  
P.R Barber ◽  
S.M Ameer-Beg ◽  
J Gilbey ◽  
L.M Carlin ◽  
M Keppler ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Protein Interactions ◽  
Time Domain ◽  
Single Photon ◽  
Global Analysis ◽  
Exponential Fitting ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Protein Protein Interactions ◽  
Lifetime Imaging

Förster resonance energy transfer (FRET) detected via fluorescence lifetime imaging microscopy (FLIM) and global analysis provide a way in which protein–protein interactions may be spatially localized and quantified within biological cells. The FRET efficiency and proportion of interacting molecules have been determined using bi-exponential fitting to time-domain FLIM data from a multiphoton time-correlated single-photon counting microscope system. The analysis has been made more robust to noise and significantly faster using global fitting, allowing higher spatial resolutions and/or lower acquisition times. Data have been simulated, as well as acquired from cell experiments, and the accuracy of a modified Levenberg–Marquardt fitting technique has been explored. Multi-image global analysis has been used to follow the epidermal growth factor-induced activation of Cdc42 in a short-image-interval time-lapse FLIM/FRET experiment. Our implementation offers practical analysis and time-resolved-image manipulation, which have been targeted towards providing fast execution, robustness to low photon counts, quantitative results and amenability to automation and batch processing.

scholarly journals Tracking metabolic dynamics of apoptosis with high-speed two-photon fluorescence lifetime imaging microscopy

2019 ◽  
Vol 10 (12) ◽  
pp. 6408 ◽  
Author(s):  
Andrew J. Bower ◽  
Janet E. Sorrells ◽  
Joanne Li ◽  
Marina Marjanovic ◽  
Ronit Barkalifa ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
High Speed ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Two Photon ◽  
Lifetime Imaging ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence
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