Fluorescence lifetime imaging of molecular rotors to map microviscosity in cells (Invited Paper)

2010 ◽  
Vol 8 (10) ◽  
pp. 926-930 ◽  
Author(s):  
James A. Levitt James A. Levitt ◽  
Marina K. Kuimova Marina K. Kuimova ◽  
Gokhan Yahioglu Gokhan Yahioglu ◽  
Pei-Hua Chung Pei-Hua Chung ◽  
Klaus Suhling Klaus Suhling ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Molecular Rotors ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
In Cells

scholarly journals Systematic Enzyme Mapping of Cellular Metabolism by Phasor-Analyzed Label-Free NAD(P)H Fluorescence Lifetime Imaging

2019 ◽  
Vol 20 (22) ◽  
pp. 5565 ◽  
Author(s):  
Leben ◽  
Köhler ◽  
Radbruch ◽  
Hauser ◽  
Niesner
Keyword(s):  
Fluorescence Lifetime ◽  
Metabolic Activity ◽  
Cellular Metabolism ◽  
Fluorescence Lifetime Imaging ◽  
Label Free ◽  
Analysis Framework ◽  
Cell Nuclei ◽  
Lifetime Imaging ◽  
Subcellular Resolution ◽  
In Cells

In the past years, cellular metabolism of the immune system experienced a revival, as it has become clear that it is not merely responsible for the cellular energy supply, but also impacts on many signaling pathways and, thus, on diverse cellular functions. Label-free fluorescence lifetime imaging of the ubiquitous coenzymes NADH and NADPH (NAD(P)H-FLIM) makes it possible to monitor cellular metabolism in living cells and tissues and has already been applied to study metabolic changes both under physiologic and pathologic conditions. However, due to the complex distribution of NAD(P)H-dependent enzymes in cells, whose distribution continuously changes over time, a thorough interpretation of NAD(P)H-FLIM results, in particular, resolving the contribution of various enzymes to the overall metabolic activity, remains challenging. We developed a systematic framework based on angle similarities of the phase vectors and their length to analyze NAD(P)H-FLIM data of cells and tissues based on a generally valid reference system of highly abundant NAD(P)H-dependent enzymes in cells. By using our analysis framework, we retrieve information not only about the overall metabolic activity, i.e., the fraction of free to enzyme-bound NAD(P)H, but also identified the enzymes predominantly active within the sample at a certain time point with subcellular resolution. We verified the performance of the approach by applying NAD(P)H-FLIM on a stromal-like cell line and identified a different group of enzymes that were active in the cell nuclei as compared to the cytoplasm. As the systematic phasor-based analysis framework of label-free NAD(P)H-FLIM can be applied both in vitro and in vivo, it retains the unique power to enable dynamic enzyme-based metabolic investigations, at subcellular resolution, in genuine environments.

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 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

scholarly journals Monitoring conformational changes of proteins in cells by fluorescence lifetime imaging microscopy

2003 ◽  
Vol 373 (3) ◽  
pp. 999-999
Author(s):  
V. CALLEJA ◽  
S. M. AMEER-BEG ◽  
B. VOJNOVIC ◽  
R. WOSCHOLSKI ◽  
J. DOWNWARD ◽  
...  
Keyword(s):  
Conformational Changes ◽  
Fluorescence Lifetime ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
In Cells
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