Towards in-vivo assessment of fluorescence lifetime: imaging using time-gated intensified CCD camera

2018 ◽  
Vol 38 (4) ◽  
pp. 966-974 ◽  
Author(s):  
Piotr Sawosz ◽  
Stanislaw Wojtkiewicz ◽  
Michal Kacprzak ◽  
Elzbieta Zieminska ◽  
Magdalena Morawiec ◽  
...  
2014 ◽  
Vol 20 (13) ◽  
pp. 3531-3539 ◽  
Author(s):  
Yasaman Ardeshirpour ◽  
Victor Chernomordik ◽  
Moinuddin Hassan ◽  
Rafal Zielinski ◽  
Jacek Capala ◽  
...  

2020 ◽  
Author(s):  
Xingbo Yang ◽  
Daniel J. Needleman

AbstractMitochondria are central to metabolism and their dysfunctions are associated with many diseases1–9. Metabolic flux, the rate of turnover of molecules through a metabolic pathway, is one of the most important quantities in metabolism, but it remains a challenge to measure spatiotemporal variations in mitochondrial metabolic fluxes in living cells. Fluorescence lifetime imaging microscopy (FLIM) of NADH is a label-free technique that is widely used to characterize the metabolic state of mitochondria in vivo10–18. However, the utility of this technique has been limited by the inability to relate FLIM measurement to the underlying metabolic activities in mitochondria. Here we show that, if properly interpreted, FLIM of NADH can be used to quantitatively measure the flux through a major mitochondrial metabolic pathway, the electron transport chain (ETC), in vivo with subcellular resolution. This result is based on the use of a coarse-grained NADH redox model, which we test in mouse oocytes subject to a wide variety of perturbations by comparing predicted fluxes to direct biochemical measurements and by self-consistency criterion. Using this method, we discovered a subcellular spatial gradient of mitochondrial metabolic flux in mouse oocytes. We showed that this subcellular variation in mitochondrial flux correlates with a corresponding subcellular variation in mitochondrial membrane potential. The developed model, and the resulting procedure for analyzing FLIM of NADH, are valid under nearly all circumstances of biological interest. Thus, this approach is a general procedure to measure metabolic fluxes dynamically in living cells, with subcellular resolution.


2020 ◽  
Vol 8 (3) ◽  
pp. 034003
Author(s):  
Deborah S Barkauskas ◽  
Gregory Medley ◽  
Xiaowen Liang ◽  
Yousuf H Mohammed ◽  
Camilla A Thorling ◽  
...  

2019 ◽  
Vol 10 (15) ◽  
pp. 4227-4235 ◽  
Author(s):  
Yingying Ning ◽  
Shengming Cheng ◽  
Jing-Xiang Wang ◽  
Yi-Wei Liu ◽  
Wei Feng ◽  
...  

Lanthanide complex was successfully applied in the design of pH-responsive NIR τ probe for quantitative in vivo imaging.


2004 ◽  
Vol 34 (1) ◽  
pp. 19-27 ◽  
Author(s):  
Shu Zhang ◽  
Guozheng Wang ◽  
David G. Fernig ◽  
Philip S. Rudland ◽  
Stephen E. D. Webb ◽  
...  

2007 ◽  
Vol 6 (5) ◽  
pp. 7290.2007.00030 ◽  
Author(s):  
Abedelnasser Abulrob ◽  
Eric Brunette ◽  
Jacqueline Slinn ◽  
Ewa Baumann ◽  
Danica Stanimirovic

Fluorescence lifetime is an intrinsic parameter of the fluorescent probe, independent of the probe concentration but sensitive to changes in the surrounding microenvironment. Therefore, fluorescence lifetime imaging could potentially be applied to in vivo diagnostic assessment of changes in the tissue microenvironment caused by disease, such as ischemia. The aim of this study was to evaluate the utility of noninvasive fluorescence lifetime imaging in distinguishing between normal and ischemic kidney tissue in vivo. Mice were subjected to 60-minute unilateral kidney ischemia followed by 6-hour reperfusion. Animals were then injected with the near-infrared fluorescence probe Cy5.5 or saline and imaged using a time-domain small-animal optical imaging system. Both fluorescence intensity and lifetime were acquired. The fluorescence intensity of Cy5.5 was clearly reduced in the ischemic compared with the contralateral kidney, and the fluorescence lifetime of Cy5.5 was not detected in the ischemic kidney, suggesting reduced kidney clearance. Interestingly, the two-component lifetime analysis of endogenous fluorescence at 700 nm distinguished renal ischemia in vivo without the need for Cy5.5 injection for contrast enhancement. The average fluorescence lifetime of endogenous tissue fluorophores was a sensitive indicator of kidney ischemia ex vivo. The study suggests that fluorescence lifetime analysis of endogenous tissue fluorophores could be used to discriminate ischemic or necrotic tissues by noninvasive in vivo or ex vivo organ imaging.


2016 ◽  
Vol 127 (3) ◽  
pp. 473-482 ◽  
Author(s):  
Sven R. Kantelhardt ◽  
Darius Kalasauskas ◽  
Karsten König ◽  
Ella Kim ◽  
Martin Weinigel ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document