Measuring macromolecular crowding in cells through fluorescence anisotropy imaging with an AIE fluorogen

Soleimaninejad, Hamid; Chen, Moore Z.; Lou, Xiaoding; Smith, Trevor A.; Hong, Yuning

doi:10.1039/c6cc09916e

Cited by 43 publications

(25 citation statements)

References 29 publications

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“…1, G is a correction D r a f t factor (0.98) to account for any polarization bias of the detection system and B is a background term, which is subtracted from each image using ImageJ. [22][23] (1) = Defocusing was achieved by manually moving the microscope objective away from the sample by ~ 1 micron. The emitted light was collected through the objective, separated from the excitation light by a dichroic mirror (Chroma Technology) and a further long pass filter (Chroma Technology) then passed through a 3.3  beam expander (Olympus) onto the CCD camera.…”

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confidence: 99%

Fluorescence anisotropy imaging of a polydiacetylene photopolymer film

et al. 2019

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UV-illumination of phase-separated surfactant films prepared from mixtures of photopolymerizable 10,12-pentacosadiynoic acid and perfluorotetradecanoic acid results in the formation of fluorescent polydiacetylene fibers and aggregates. In this work, the orientation of polymer strands that comprise the resulting photopolymer structures has been probed using fluorescence anisotropy imaging in combination with defocused single-molecule fluorescence imaging. Imaging experiments indicate the presence of significant fiber-to-fiber heterogeneity, as well as anisotropy within each fiber (or aggregate), with both of these properties changing as a function of film preparation conditions. This anisotropy can be attributed to various alignments of the constituent polymer strands that comprise the larger fibers and aggregates. Intriguingly, when using defocused imaging, fiber images consisted of a series of discrete “doughnut” fluorescence emission patterns, which exhibited intermittent on–off blinking behavior; both of these properties are characteristic of individual emission transition dipoles (single molecules). Further, all of the individual emission transition dipoles had a uniform orientation with respect to the axis of the fiber, indicating a common orientation of discrete emitters in the larger polymer fiber. The implications of these results for future studies of the electronic properties of conjugated polymers in larger macroscopic systems are noted.

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confidence: 99%

Fluorescence anisotropy imaging of a polydiacetylene photopolymer film

et al. 2019

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“…In contrast to the method of using delicate FRET probes, Soleimaninejad and co‐workers reported a simple AIEgen, TPE‐Py‐NCS, for measuring the same macromolecular crowding environment in cells (Figure ) . The amine‐reactive AIEgen was first incubated with living cells to label cytosolic proteins through the amine‐mediated reactions.…”

Section: Probing Polymer Conformational Transitions Of Proteinsmentioning

confidence: 99%

“…In contrastt ot he method of using delicate FRET probes, [72] Soleimaninejad and co-workersr eported as imple AIEgen, TPE-Py-NCS, for measuring the same macromolecular crowding environmenti nc ells ( Figure 14). [73] The amine-reactive AIEgen was first incubated with living cells to label cytosolic proteins through the amine-mediated reactions. Because the AIEgen is highly flexible in structure, rotational freedom would be sensitive to changes to the crowding conditions, and could be studied by means of FAIM techniques.…”

Section: Protein Crowdingmentioning

confidence: 99%

Fluorogenic Detection and Characterization of Proteins by Aggregation‐Induced Emission Methods

Xie

Wong

Chen

et al. 2019

Chemistry A European J

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Protein is one of the four most important biomacromolecules in living systems. The detection, quantification, localization, and characterization of proteins is essential for an understanding of biological fundamentals, as well as for the diagnostics and treatment of protein‐related diseases. By using intrinsic and extrinsic fluorescence, different techniques have been established to study proteins, many of which are now being routinely used in research laboratories and clinics. This review summarizes the applications of aggregation‐induced emission (AIE) fluorescence in protein science. In contrast to traditional fluorescent dyes, the activation of AIE dyes is mainly attributed to the restriction of intramolecular motions. This unique turn‐on mechanism of AIE dyes allows researchers to develop novel fluorogenic strategies for sensitive, selective, and reliable analysis of proteins. This review focuses on introducing AIE strategies for 1) detection, localization, and quantification of proteins; 2) probing polymer conformational transitions of proteins; 3) characterization of protein–ligand interactions; and 4) evaluation of enzyme activities. Perspectives and challenges with respect to this emerging field of protein characterization are offered.

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“…Symbolic examples of AIE-active dyes include a series of tetraphenylethene (TPE) derivatives comprised of an olefin stator surrounded by four phenyl rotators. They have been proved valuable in visualization of biomolecules and cellular environments such as mitochondrial phospholipids [ 8 ], nucleic acids [ 9 ], tumor proteins [ 10 ], and changes in intracellular pH [ 11 ], viscosity [ 12 ] and macromolecular crowding conditions [ 13 ]. The distyrylanthracene (DSA) derivatives are another set of typical AIE-active dyes ( Figure 1 left).…”

Section: Introductionmentioning

confidence: 99%

9-Vinylanthracene Based Fluorogens: Synthesis, Structure-Property Relationships and Applications

et al. 2017

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Fluorescent dyes with aggregation-induced emission (AIE) properties exhibit intensified emission upon aggregation. They are promising candidates to study biomolecules and cellular changes in aqueous environments when aggregation formation occurs. Here, we report a group of 9-position functionalized anthracene derivatives that were conveniently synthesized by the palladium-catalyzed Heck reaction. Using fluorometric analyses, these dyes were confirmed to show AIE behavior upon forming aggregates at high concentrations, in viscous solvents, and when poorly solubilized. Their photophysical properties were then further correlated with their structural features, using density functional theory (DFT) calculation. Finally, we demonstrated their potential applications in monitoring pH changes, quantifying globular proteins, as well as cell imaging with confocal microscopy.

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Measuring macromolecular crowding in cells through fluorescence anisotropy imaging with an AIE fluorogen

Cited by 43 publications

References 29 publications

Fluorescence anisotropy imaging of a polydiacetylene photopolymer film

Fluorescence anisotropy imaging of a polydiacetylene photopolymer film

Fluorogenic Detection and Characterization of Proteins by Aggregation‐Induced Emission Methods

9-Vinylanthracene Based Fluorogens: Synthesis, Structure-Property Relationships and Applications

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