Fluorescence Lifetime Imaging Microscopy

Chang, Ching Wei; Sud, Dhruv; Mycek, Mary Ann

doi:10.1016/s0091-679x(06)81024-1

Cited by 143 publications

(121 citation statements)

References 44 publications

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“…Tests for significant differences from control were performed by one-way ANOVA. Fluorescence lifetime FRET measurements have certain advantages over intensity-based methods, because fluorescence lifetime is an intrinsic property of the fluorescent molecule that is not influenced by factors such as excitation source, detection gain, optical loss, and variation in fluorophore concentration and is much less sensitive to variation in the donor-acceptor ratio (assuming an excess of acceptor) (16).…”

Section: Methodsmentioning

confidence: 99%

Nonsteroidal Anti-inflammatory Drugs Alter the Spatiotemporal Organization of Ras Proteins on the Plasma Membrane

Zhou

Cho

Plowman

et al. 2012

Journal of Biological Chemistry

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Background: Ras proteins form structurally distinct nanoclusters on cell plasma membrane that are critical for signal transduction. Results: Amphiphilic anti-inflammatory agents stabilize cholesterol-dependent Ras nanoclusters, perturb GTP-dependent lateral segregation, and compromise nanocluster separation. Conclusion: Biological amphiphiles have wide-ranging effects on plasma membrane heterogeneity and protein nanoclustering. Significance: This is a novel mechanism for altering membrane protein segregation that has important consequences for cell signaling.

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Section: Methodsmentioning

confidence: 99%

Nonsteroidal Anti-inflammatory Drugs Alter the Spatiotemporal Organization of Ras Proteins on the Plasma Membrane

Zhou

Cho

Plowman

et al. 2012

Journal of Biological Chemistry

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“…Consistent with our previous studies, SF irradiation led to retraction of the severed ends of the targeted SF (top row), and inspection of the donor and FRET intensity (middle rows) revealed that the VinTS localizes to FAs as expected. Because the FRET intensity also depends on factors unrelated to donoracceptor separation (Chang and Mycek, 2012;Chang et al, 2007;Chang et al, 2009;Zhong et al, 2007), such as the amount of localized VinTS, we created FRET ratio maps in which we masked the FRET intensity to show only FAs and normalized this intensity by the donor intensity (bottom row). In these maps, an increasing FRET ratio corresponds to an increasing FRET signal (actual degree of energy transfer) or decreasing tension, which can in turn be tracked on an FA-by-FA basis.…”

Section: Mapping Tension Distributions Of Single Stress Fibersmentioning

confidence: 99%

Vinculin tension distributions of individual stress fibers within cell-matrix adhesions

Chang

Kumar

2013

Journal of Cell Science

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SummaryActomyosin stress fibers (SFs) enable cells to exert traction on planar extracellular matrices (ECMs) by tensing focal adhesions (FAs) at the cell-ECM interface. Although it is widely appreciated that the spatial and temporal distribution of these tensile forces play key roles in polarity, motility, fate choice, and other defining cell behaviors, virtually nothing is known about how an individual SF quantitatively contributes to tensile loads borne by specific molecules within associated FAs. We address this key open question by using femtosecond laser ablation to sever single SFs in cells while tracking tension across vinculin using a molecular optical sensor. We show that disruption of a single SF reduces tension across vinculin in FAs located throughout the cell, with enriched vinculin tension reduction in FAs oriented parallel to the targeted SF. Remarkably, however, some subpopulations of FAs exhibit enhanced vinculin tension upon SF irradiation and undergo dramatic, unexpected transitions between tension-enhanced and tension-reduced states. These changes depend strongly on the location of the severed SF, consistent with our earlier finding that different SF pools are regulated by distinct myosin activators. We critically discuss the extent to which these measurements can be interpreted in terms of whole-FA tension and traction and propose a model that relates SF tension to adhesive loads and cell shape stability. These studies represent the most direct and highresolution intracellular measurements of SF contributions to tension on specific FA proteins to date and offer a new paradigm for investigating regulation of adhesive complexes by cytoskeletal force.

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“…As there is a decrease in the dot size there will an increase in the emitted photon energy. Phase separation is used for forming large area of mono layers of quantum dots [7,19]. …”

Section: Introductionmentioning

confidence: 99%

Quantum dots wide absorption pattern, photo stability and greater quantum yields provides a gate way to power full applications in nano medicine and nano biotechnology

Abilash¹

2017

Med-Science

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Quantum dots have specific optical and electronic properties which makes their wide application in the field of nano bio technology. The emitted photon energy in a quantum dot increases with a decrease in the dot size. Since quantum dots are tunable hence by injecting quantum dot based probes multi color imaging of tumor cells, cancer cells and lymph nodes can be accomplished which is very advantageous in the medical field. Various researches have been done and much is going on where quantum dots works as an efficient drug carrier system during analysis or treatment in targeted drug delivery. Quantum dots can be applied to cell for drug screening and analysis. Linking with antibodies as biological labels makes them appropriate for various applications in cellular and molecular imaging.

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Fluorescence Lifetime Imaging Microscopy

Cited by 143 publications

References 44 publications

Nonsteroidal Anti-inflammatory Drugs Alter the Spatiotemporal Organization of Ras Proteins on the Plasma Membrane

Nonsteroidal Anti-inflammatory Drugs Alter the Spatiotemporal Organization of Ras Proteins on the Plasma Membrane

Vinculin tension distributions of individual stress fibers within cell-matrix adhesions

Quantum dots wide absorption pattern, photo stability and greater quantum yields provides a gate way to power full applications in nano medicine and nano biotechnology

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