scholarly journals Membrane-Bound Molecular Rotors Measure Viscosity in Live Cells via Fluorescence Lifetime Imaging

2009 ◽  
Vol 113 (27) ◽  
pp. 11634-11642 ◽  
Author(s):  
James A. Levitt ◽  
Marina K. Kuimova ◽  
Gokhan Yahioglu ◽  
Pei-Hua Chung ◽  
Klaus Suhling ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Molecular Rotors ◽  
Live Cells ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Membrane Bound

scholarly journals Streamlined histone-based fluorescence lifetime imaging microscopy reveals ATM regulation of chromatin compaction

2017 ◽  
Author(s):  
Alice Sherrard ◽  
Paul Bishop ◽  
Melanie Panagi ◽  
Maria Beatriz Villagomez ◽  
Dominic Alibhai ◽  
...  
Keyword(s):  
Dna Damage ◽  
Fluorescence Lifetime ◽  
Genotoxic Stress ◽  
Live Cells ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Chromatin Compaction ◽  
Lifetime Imaging ◽  
Detailed Method ◽  
Chromatin Organisation

AbstractChanges in chromatin compaction are crucial during genomic responses. Thus, methods that enable such measurements are instrumental for investigating genome function. Here, we address this challenge by developing, validating, and streamlining histone-based fluorescence lifetime imaging microscopy (FLIM) that robustly detects chromatin compaction states in fixed and live cells; in 2D and 3D. We present quality-controlled and detailed method that is simpler and faster than previous approches, and uses FLIMfit open-source software. We demonstrate the versatility of our method through its combination with immunofluorescence and its implementation in immortalised cells and primary neurons. Owing to these developments, we applied this method to elucidate the function of the DNA damage response kinase, ATM, in regulating chromatin organisation after genotoxic-stress. We unravelled a role for ATM in regulating chromatin compaction independently of DNA damage. Collectively, we present an adaptable chromatin FLIM method for examining chromatin structure in cells, and establish its broader utility.

Spectroscopy and fluorescence lifetime imaging in live cells of a cyano-substituted combretastatin

2014 ◽  
Vol 3 (3) ◽  
pp. 211-218 ◽  
Author(s):  
Kathrin M. Scherer ◽  
Roger H. Bisby ◽  
Stanley W. Botchway ◽  
Greg M. Greetham ◽  
John A. Hadfield ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Live Cells ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

scholarly journals Quantitative sensing of microviscosity in protocells and amyloid materials using fluorescence lifetime imaging of molecular rotors

2014 ◽  
Author(s):  
Alex J. Thompson ◽  
T.-Y. Dora Tang ◽  
Therese W. Herling ◽  
C. Rohaida Che Hak ◽  
Stephen Mann ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Molecular Rotors ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

scholarly journals A novel technique for mapping viscosity in discrete subcellular locations with a BODIPY based fluorescent probe

2019 ◽  
Author(s):  
Lior Pytowski ◽  
Alex C. Foley ◽  
Zayra E. Hernández ◽  
Niall Moon ◽  
Tim Donohoe ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging ◽  
Mitochondrial Matrix ◽  
Molecular Rotor ◽  
Cellular Processes ◽  
Lifetime Imaging ◽  
Novel Technique ◽  
Absolute Viscosity ◽  
Membrane Bound ◽  
Fluorescent Molecules

AbstractNumerous cellular processes, including enzyme behaviour, signalling, and protein folding and transport are highly influenced by the local microviscosity environment within living cells. Molecular rotors are fluorescent molecules that respond to the viscosity of their environment through changes in both the intensity and lifetime of their fluorescence. We have synthesised a novel boron-dipyrrin (BODIPY) molecular rotor that is also a substrate for the SNAP-tag targeting system (named BG-BODIPY), allowing us to target the rotor to discrete locations within the living cell. We demonstrate that BG-BODIPY reports viscosity, and that this can be measured either through fluorescence lifetime or intensity ratiometric measurements. The relative microviscosities within the ER, Golgi, mitochondrial matrix, peroxisomes, lysosomes, cytoplasm, and nucleoplasm were significantly different. Additionally, this approach permitted fluorescence lifetime imaging microscopy (FLIM) to determine the absolute viscosity within both mitochondria and stress granules, showcasing BG-BODIPY’s usefulness in studying both membrane bound and membraneless organelles. These results highlight targeted BG-BODIPY’s broad usefulness for making measurements of cellular viscosity both with FLIM and conventional confocal microscopy, the latter option greatly extending the accessibility of the technique.

scholarly journals Multiplexable fluorescence lifetime imaging (FLIM) probes for Abl and Src-family kinases

2020 ◽  
Vol 56 (87) ◽  
pp. 13409-13412
Author(s):  
Sampreeti Jena ◽  
Nur P. Damayanti ◽  
Jackie Tan ◽  
Erica D. Pratt ◽  
Joseph M. K. Irudayaraj ◽  
...  
Keyword(s):  
Single Cell ◽  
Fluorescence Lifetime ◽  
Src Family Kinases ◽  
Live Cells ◽  
Cell Penetrating Peptide ◽  
Fluorescence Lifetime Imaging ◽  
Src Family Kinase ◽  
Peptide Substrates ◽  
Lifetime Imaging ◽  
Cell Penetrating

Here we describe development of cell-penetrating peptide substrates for fluorescence lifetime imaging (FLIM) of Abl and Src-family kinase activities, applied to mapping single cell and subcellular dynamics of multiple kinases in live cells.

scholarly journals Mapping microbubble viscosity using fluorescence lifetime imaging of molecular rotors

2013 ◽  
Vol 110 (23) ◽  
pp. 9225-9230 ◽  
Author(s):  
Neveen A. Hosny ◽  
Graciela Mohamedi ◽  
Paul Rademeyer ◽  
Joshua Owen ◽  
Yilei Wu ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Molecular Rotors ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging
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