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      Quantitative Investigation for the Dielectrophoretic Effect of Fluorescent Dyes at Single-Cell Resolution

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          Abstract

          Most of the microscopy-based, quantitative assays rely on fluorescent dyes. In this study, we investigated the impact of fluorescent dyes on the dielectrophoretic response of the mammalian cells. The dielectrophoretic measurements were performed to quantify whether the fluorescent dyes alter the dielectrophoretic properties of the cells at single-cell resolution. Our results present that when 10 V pp electric field is applied, the fluorescent-labeled cells experienced the crossover frequency at 8–10 kHz, whereas the label-free cells exhibited at 16–18 kHz.

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          The Motion and Precipitation of Suspensoids in Divergent Electric Fields

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            Fluorescent dyes and their supramolecular host/guest complexes with macrocycles in aqueous solution.

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              Fluorescent proteins for live-cell imaging with super-resolution.

              Fluorescent proteins (FPs) from the GFP family have become indispensable as marker tools for imaging live cells, tissues and entire organisms. A wide variety of these proteins have been isolated from natural sources and engineered to optimize their properties as genetically encoded markers. Here we review recent developments in this field. A special focus is placed on photoactivatable FPs, for which the fluorescence emission can be controlled by light irradiation at specific wavelengths. They enable regional optical marking in pulse-chase experiments on live cells and tissues, and they are essential marker tools for live-cell optical imaging with super-resolution. Photoconvertible FPs, which can be activated irreversibly via a photo-induced chemical reaction that either turns on their emission or changes their emission wavelength, are excellent markers for localization-based super-resolution microscopy (e.g., PALM). Patterned illumination microscopy (e.g., RESOLFT), however, requires markers that can be reversibly photoactivated many times. Photoswitchable FPs can be toggled repeatedly between a fluorescent and a non-fluorescent state by means of a light-induced chromophore isomerization coupled to a protonation reaction. We discuss the mechanistic origins of the effect and illustrate how photoswitchable FPs are employed in RESOLFT imaging. For this purpose, special FP variants with low switching fatigue have been introduced in recent years. Despite nearly two decades of FP engineering by many laboratories, there is still room for further improvement of these important markers for live cell imaging.
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                Author and article information

                Journal
                ACS Omega
                ACS Omega
                ao
                acsodf
                ACS Omega
                American Chemical Society
                2470-1343
                03 July 2018
                31 July 2018
                : 3
                : 7
                : 7243-7246
                Affiliations
                []Faculty of Engineering and Natural Sciences, Sabanci University , Istanbul 34956, Turkey
                []Mechanical Engineering Department, Clemson University , Clemson, South Carolina 29634-0921, United States
                Author notes
                Article
                10.1021/acsomega.8b00541
                6068594
                ebe32997-a0a5-4312-a3e1-7eafb8a1934f
                Copyright © 2018 American Chemical Society

                This is an open access article published under an ACS AuthorChoice License, which permits copying and redistribution of the article or any adaptations for non-commercial purposes.

                History
                : 21 March 2018
                : 20 June 2018
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                ao8b00541
                ao-2018-00541m

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