scholarly journals Dynamic conformational transitions of the EGF receptor in living mammalian cells determined by FRET and fluorescence lifetime imaging microscopy

2013 ◽  
Vol 83 (9) ◽  
pp. 794-805 ◽  
Author(s):  
Iwona Ziomkiewicz ◽  
Anastasia Loman ◽  
Reinhard Klement ◽  
Cornelia Fritsch ◽  
Andrey S. Klymchenko ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Mammalian Cells ◽  
Egf Receptor ◽  
Conformational Transitions ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

scholarly journals Visualising G-quadruplex DNA dynamics in live cells by fluorescence lifetime imaging microscopy

2020 ◽  
Author(s):  
Peter Andrew Summers ◽  
Ben Lewis ◽  
Jorge Gonzalez-Garcia ◽  
Rosa Maria Porreca ◽  
Aaron H M Lim ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Mammalian Cells ◽  
Live Cells ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Dna Dynamics ◽  
Drug Candidates ◽  
Lifetime Imaging ◽  
Wide Range

Guanine rich regions of oligonucleotides fold into quadruple-stranded structures called G-quadruplexes (G4). Increasing evidence suggests that these G4 structures form in vivo and play a crucial role in cellular processes. However, their direct observation in live cells remains a challenge. Here we demonstrate that a fluorescent probe (DAOTA-M2) in conjunction with Fluorescence Lifetime Imaging Microscopy (FLIM) can identify G4 within nuclei of live and fixed cells. We present a new FLIM-based cellular assay to study the interaction of non-fluorescent small molecules with G4 and apply it to a wide range of drug candidates. We also demonstrate that DAOTA-M2 can be used to study G4 stability in live cells. Reduction of FancJ and RTEL1 expression in mammalian cells increases the DAOTA-M2 lifetime and therefore suggests an increased number of G4 in these cells, implying that FancJ and RTEL1 play a role in resolving G4 structures in cellulo.

scholarly journals Visualising G-quadruplex DNA dynamics in live cells by fluorescence lifetime imaging microscopy

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Peter A. Summers ◽  
Benjamin W. Lewis ◽  
Jorge Gonzalez-Garcia ◽  
Rosa M. Porreca ◽  
Aaron H. M. Lim ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Mammalian Cells ◽  
Live Cells ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Dna Dynamics ◽  
Drug Candidates ◽  
Lifetime Imaging ◽  
Wide Range

AbstractGuanine rich regions of oligonucleotides fold into quadruple-stranded structures called G-quadruplexes (G4s). Increasing evidence suggests that these G4 structures form in vivo and play a crucial role in cellular processes. However, their direct observation in live cells remains a challenge. Here we demonstrate that a fluorescent probe (DAOTA-M2) in conjunction with fluorescence lifetime imaging microscopy (FLIM) can identify G4s within nuclei of live and fixed cells. We present a FLIM-based cellular assay to study the interaction of non-fluorescent small molecules with G4s and apply it to a wide range of drug candidates. We also demonstrate that DAOTA-M2 can be used to study G4 stability in live cells. Reduction of FancJ and RTEL1 expression in mammalian cells increases the DAOTA-M2 lifetime and therefore suggests an increased number of G4s in these cells, implying that FancJ and RTEL1 play a role in resolving G4 structures in cellulo.

scholarly journals Direct organelle thermometry with fluorescence lifetime imaging microscopy in single myotubes

2016 ◽  
Vol 52 (24) ◽  
pp. 4458-4461 ◽  
Author(s):  
Hideki Itoh ◽  
Satoshi Arai ◽  
Thankiah Sudhaharan ◽  
Sung-Chan Lee ◽  
Young-Tae Chang ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Heat Production ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
C2c12 Myotube

FLIM of ER thermo yellow and non-targeted mCherry reveals the Ca2+-dependent heat production localized to SR in C2C12 myotube.

Sign in / Sign up

Export Citation Format

Share Document