One-step synthesis of water-dispersible silicon nanoparticles and their use in fluorescence lifetime imaging of living cells

2014 ◽  
Vol 2 (27) ◽  
pp. 4338-4345 ◽  
Author(s):  
Jing Wang ◽  
Dai-Xin Ye ◽  
Guo-Hai Liang ◽  
Jian Chang ◽  
Ji-Lie Kong ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Fluorescence Intensity ◽  
Silicon Nanoparticles ◽  
Living Cells ◽  
Fluorescence Lifetime Imaging ◽  
Kb Cells ◽  
Water Dispersible ◽  
Lifetime Imaging ◽  
One Step

Fluorescence intensity (upper panel) and lifetime (bottom panel) images of KB cells stained with FITC (B), Si NPs (C) and unstained control (A).

scholarly journals Carbon Dots for Intracellular pH Sensing with Fluorescence Lifetime Imaging Microscopy

Nanomaterials ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 604 ◽  
Author(s):  
Maojia Huang ◽  
Xinyue Liang ◽  
Zixiao Zhang ◽  
Jing Wang ◽  
Yiyan Fei ◽  
...  
Keyword(s):  
Intracellular Ph ◽  
Fluorescence Lifetime ◽  
Fluorescence Intensity ◽  
Carbon Dots ◽  
Living Cells ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Gradual Change ◽  
Ph Sensing ◽  
Lifetime Imaging

The monitoring of intracellular pH is of great importance for understanding intracellular trafficking and functions. It has various limitations for biosensing based on the fluorescence intensity or spectra study. In this research, pH-sensitive carbon dots (CDs) were employed for intracellular pH sensing with fluorescence lifetime imaging microscopy (FLIM) for the first time. FLIM is a highly sensitive method that is used to detect a microenvironment and it can overcome the limitations of biosensing methods based on fluorescence intensity. The different groups on the CDs surfaces changing with pH environments led to different fluorescence lifetime values. The CDs aqueous solution had a gradual change from 1.6 ns to 3.7 ns in the fluorescence lifetime with a pH range of 2.6–8.6. Similar fluorescence lifetime changes were found in pH buffer-treated living cells. The detection of lysosomes, cytoplasm, and nuclei in living cells was achieved by measuring the fluorescence lifetime of CDs. In particular, a phasor FLIM analysis was used to improve the pH imaging. Moreover, the effects of the coenzymes, amino acids, and proteins on the fluorescence lifetime of CDs were examined in order to mimic the complex microenvironment inside the cells.

scholarly journals Coarse-grained modeling of mitochondrial metabolism enables subcellular flux inference from fluorescence lifetime imaging microscopy

2020 ◽  
Author(s):  
Xingbo Yang ◽  
Daniel J. Needleman
Keyword(s):  
Fluorescence Lifetime ◽  
Metabolic Flux ◽  
Living Cells ◽  
Coarse Grained ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Metabolic Fluxes ◽  
Subcellular Resolution

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.

Fast fluorescence lifetime imaging of calcium in living cells

2004 ◽  
Vol 9 (6) ◽  
pp. 1230 ◽  
Author(s):  
A. V. Agronskaia ◽  
L. Tertoolen ◽  
H. C. Gerritsen
Keyword(s):  
Fluorescence Lifetime ◽  
Living Cells ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging

scholarly journals A simple yet effective AIE-based fluorescent nano-thermometer for temperature mapping in living cells using fluorescence lifetime imaging microscopy

2020 ◽  
Vol 5 (3) ◽  
pp. 488-494 ◽  
Author(s):  
Hui Gao ◽  
Chuen Kam ◽  
Tsu Yu Chou ◽  
Ming-Yu Wu ◽  
Xin Zhao ◽  
...  
Keyword(s):  
Fluorescence Lifetime ◽  
Living Cells ◽  
Fluorescence Lifetime Imaging Microscopy ◽  
Fluorescence Lifetime Imaging ◽  
Temperature Mapping ◽  
Lifetime Imaging

A novel nano-thermometer composed of butter and AIE molecules can be used for intracellular temperature mapping using fluorescence lifetime imaging.

scholarly journals In Vivo Time Domain Optical Imaging of Renal Ischemia-Reperfusion Injury: Discrimination Based on Fluorescence Lifetime

2007 ◽  
Vol 6 (5) ◽  
pp. 7290.2007.00030 ◽  
Author(s):  
Abedelnasser Abulrob ◽  
Eric Brunette ◽  
Jacqueline Slinn ◽  
Ewa Baumann ◽  
Danica Stanimirovic
Keyword(s):  
Optical Imaging ◽  
Fluorescence Lifetime ◽  
Fluorescence Intensity ◽  
Time Domain ◽  
Ex Vivo ◽  
Renal Ischemia ◽  
Fluorescence Lifetime Imaging ◽  
Lifetime Imaging ◽  
Lifetime Analysis

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.

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