Methods for Imaging and Analyses of Intracellular Organelles Using Fluorescent and Luminescent Proteins

Takeuchi, Masaki; Ozawa, Toshio

doi:10.2116/analsci.23.25

Cited by 12 publications

(5 citation statements)

References 18 publications

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“…The commercial version Dendra2 (Evrogen) is the first monomeric red-to-green optical highlighter that has enjoyed widespread use as a tracking tool in live-cell imaging. 31,161,162 An optical highlighter isolated from another stony coral (Lobophyllia hemprichii), the tetrameric EosFP (named after the goddess of dawn in Greek mythology), behaves similarly to Kaede and the other variants described above. EosFP emits bright green fluorescence at 516 nm (Fig.…”

Section: Photoconvertable Fluorescent Proteinsmentioning

confidence: 95%

“…This highlighter features excitation and emission maxima for the green and red forms of 490/553 nm and 507/573 nm, respectively, and functions well in fusion tags for subcellular localization. The commercial version Dendra2 (Evrogen) is the first monomeric red-to-green optical highlighter that has enjoyed widespread use as a tracking tool in live-cell imaging 31,161,162…”

Section: Optical Highlighter Fluorescent Proteinsmentioning

confidence: 99%

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The fluorescent protein palette: tools for cellular imaging

2009

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This critical review provides an overview of the continually expanding family of fluorescent proteins (FPs) that have become essential tools for studies of cell biology and physiology. Here, we describe the characteristics of the genetically encoded fluorescent markers that now span the visible spectrum from deep blue to deep red. We identify some of the novel FPs that have unusual characteristics that make them useful reporters of the dynamic behaviors of proteins inside cells, and describe how many different optical methods can be combined with the FPs to provide quantitative measurements in living systems."If wood is rubbed with the Pulmo marinus, it will have all the appearance of being on fire; so much so, indeed, that a walking-stick, thus treated, will light the way like a torch" (translation of Pliny the Elder from John Bostock, 1855).

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Section: Photoconvertable Fluorescent Proteinsmentioning

confidence: 95%

Section: Optical Highlighter Fluorescent Proteinsmentioning

confidence: 99%

The fluorescent protein palette: tools for cellular imaging

2009

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“…With increased frequency of CIN induced in inflammation by cell-in-cell structure formation, we speculate that cell-in-cell structure formation might become a ‘fast track' from inflammation toward cancer transformation. 43 New in vivo cell-labeling techniques, high-resolution fluorescence imaging, nano-scale imaging techniques, tracking technologies and animal models utilizing chemically induced inflammation toward cancer 57, 58, 59 will be used to reveal the exact roles played by cell-in-cell structure formation during tumorigenesis. CIN caused by cell-in-cell structure formation is depicted in Figure 2.…”

Section: Cell-in-cell Structure Formation: An In Situ Activity or A Hmentioning

confidence: 99%

Modeling cell-in-cell structure into its biological significance

Wang

Wang³

et al. 2013

Cell Death Dis

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Although cell-in-cell structure was noted 100 years ago, the molecular mechanisms of ‘entering' and the destination of cell-in-cell remain largely unclear. It takes place among the same type of cells (homotypic cell-in-cell) or different types of cells (heterotypic cell-in-cell). Cell-in-cell formation affects both effector cells and their host cells in multiple aspects, while cell-in-cell death is under more intensive investigation. Given that cell-in-cell has an important role in maintaining homeostasis, aberrant cell-in-cell process contributes to the etiopathology in humans. Indeed, cell-in-cell is observed in many pathological processes of human diseases. In this review, we intend to discuss the biological models of cell-in-cell structures under physiological and pathological status.

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“…Fluorescent fusion proteins (FFPs), which combine FPs and proteins (either full length or portions that include organelle-targeting sequences), are widely used as genetically encoded organelle markers. Such FFPs enable us to visualize time-dependent changes in morphology, subcellular localization, and function in organelles of interest (Chudakov et al, 2010;Rizzuto et al, 1995;Takeuchi and Ozawa, 2007). For instance, the real-time tracking of early, late, and recycling endosomes with FFPs revealed that early endosomes are comprised of two dynamically distinct populations, and such differences are crucial for determining the fate of endocytic cargos in terms of undergoing degradation or recycling (Lakadamyali et al, 2006).…”

Section: /27mentioning

confidence: 99%

Folding Latency of Fluorescent Proteins Affects the Mitochondrial Localization of Fusion Proteins

Kashiwagi

Fujioka

Satoh

et al. 2019

Cell Struct. Funct.

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The discovery of fluorescent proteins (FPs) has revolutionized cell biology. The fusion of targeting sequences to FPs enables the investigation of cellular organelles and their dynamics; however, occasionally, such fluorescent fusion proteins (FFPs) exhibit behavior different from that of the native proteins. Here, we constructed a color pallet comprising different organelle markers and found that FFPs targeted to the mitochondria were mislocalized when fused to certain types of FPs. Such FPs included several variants of Aequorea victoria green FP (avGFP) and a monomeric variant of the red FP. Because the FFPs that are mislocalized include FPs with faster maturing or folding mutations, the increase in the maturation rate is likely to prevent their expected localization. Indeed, when we reintroduced amino acid substitutions so that the FP sequences were equivalent to that of wild-type avGFP, FFP localization to the mitochondria was significantly enhanced. Moreover, similar amino acid substitutions improved the localization of mitochondria-targeted pHluorin, which is a pHsensitive variant of GFP, and its capability to monitor pH changes in the mitochondrial matrix. Our findings demonstrate the importance of selecting FPs that maximize FFP function.

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Methods for Imaging and Analyses of Intracellular Organelles Using Fluorescent and Luminescent Proteins

Cited by 12 publications

References 18 publications

The fluorescent protein palette: tools for cellular imaging

The fluorescent protein palette: tools for cellular imaging

Modeling cell-in-cell structure into its biological significance

Folding Latency of Fluorescent Proteins Affects the Mitochondrial Localization of Fusion Proteins

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