Thermal Effect on Aequorea Green Fluorescent Protein Anionic and Neutral Chromophore Forms Fluorescence

Santos, Andrea M.

doi:10.1007/s10895-011-0941-0

Cited by 9 publications

(8 citation statements)

References 25 publications

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“…Temperature dependence of the fluorescence intensity of the emGFP-Mito is inverse compared to wt-GFP. In the case of emGFP-Mito increased temperature induces reduction of the hydrogen-bond network resulting in a decrease of fluorescence intensity as has been observed elsewhere 48 . The fluorescence intensity images of HeLa cells expressed with emGFP-Mito at 23–39 °C temperature is shown in Fig.…”

Section: Resultssupporting

confidence: 63%

“…1a. This spectroscopic feature was reported in the Aequorea GFP and attributed to a dipolar moment variation, resulting from a charge density transfer occurring between the close neighbors oxygen atoms 48 . Dipolar change reorganizes molecules around the electronic excited state decreasing its energy leading to observed red shift of emission maximum 49 .…”

Section: Resultsmentioning

confidence: 56%

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GFP fluorescence peak fraction analysis based nanothermometer for the assessment of exothermal mitochondria activity in live cells

Savchuk

Silvestre

Adão

et al. 2019

Sci Rep

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Nanothermometry methods with intracellular sensitivities have the potential to make important contributions to fundamental cell biology and medical fields, as temperature is a relevant physical parameter for molecular reactions to occur inside the cells and changes of local temperature are well identified therapeutic strategies. Here we show how the GFP can be used to assess temperature-based on a novel fluorescence peak fraction method. Further, we use standard GFP transfection reagents to assess temperature intracellularly in HeLa cells expressing GFP in the mitochondria. High thermal resolution and sensitivity of around 0.26% °C −1 and 2.5% °C −1 , were achieved for wt-GFP in solution and emGFP-Mito within the cell, respectively. We demonstrate that the GFP-based nanothermometer is suited to directly follow the temperature changes induced by a chemical uncoupler reagent that acts on the mitochondria. The spatial resolution allows distinguishing local heating variations within the different cellular compartments. Our discovery may lead to establishing intracellular nanothermometry as a standard method applicable to the wide range of live cells able to express GFP.

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Section: Resultssupporting

confidence: 63%

Section: Resultsmentioning

confidence: 56%

GFP fluorescence peak fraction analysis based nanothermometer for the assessment of exothermal mitochondria activity in live cells

Savchuk

Silvestre

Adão

et al. 2019

Sci Rep

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“…As demonstrated above, a low temperature during the clearing procedure facilitated the preservation of EGFP fluorescence. Previous studies have indicated that the fluorescence intensity of EGFP is also thermally sensitive ( 46 , 47 ). A higher temperature can increase the probability of GFP unfolding; moreover, the thermally induced enhancement of collisions between molecules would probably increase fluorescence quenching.…”

Section: Discussionmentioning

confidence: 99%

FDISCO: Advanced solvent-based clearing method for imaging whole organs

et al. 2019

Sci. Adv.

185

202

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“…The quenching of GFP is also significantly affected by temperature (dos Santos 2012, Schwarz et al 2015. Incubation at elevated temperature may denature GFP proteins while promoting tissue clearing.…”

Section: Preservation Of Fluorescent Signals From Reporter Proteinsmentioning

confidence: 99%

Chemical Principles in Tissue Clearing and Staining Protocols for Whole-Body Cell Profiling

Tainaka

Kuno

Kubota

et al. 2016

Annu. Rev. Cell Dev. Biol.

226

239

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Mammalian bodies have more than a billion cells per cubic centimeter, which makes whole-body cell (WBC) profiling of an organism one of the ultimate challenges in biology and medicine. Recent advances in tissue-clearing technology have enabled rapid and comprehensive cellular analyses in whole organs and in the whole body by a combination of state-of-the-art technologies of optical imaging and image informatics. In this review, we focus mainly on the chemical principles in currently available techniques for tissue clearing and staining to facilitate our understanding of their underlying mechanisms. Tissue clearing is usually conducted by the following steps: (a) fixation, (b) permeabilization, (c) decolorizing, and (d) refractive index (RI) matching. To phenotype individual cells after tissue clearing, it is important to visualize genetically encoded fluorescent reporters and/or to stain tissues with fluorescent dyes, fluorescent labeled antibodies, or nucleic acid probes. Although some technical challenges remain, the chemical principles in tissue clearing and staining for WBC profiling will enable various applications, such as identifying cellular circuits across multiple organs and measuring their dynamics in stochastic and proliferative cellular processes, for example, autoimmune and malignant neoplastic diseases.

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Thermal Effect on Aequorea Green Fluorescent Protein Anionic and Neutral Chromophore Forms Fluorescence

Cited by 9 publications

References 25 publications

GFP fluorescence peak fraction analysis based nanothermometer for the assessment of exothermal mitochondria activity in live cells

GFP fluorescence peak fraction analysis based nanothermometer for the assessment of exothermal mitochondria activity in live cells

FDISCO: Advanced solvent-based clearing method for imaging whole organs

Chemical Principles in Tissue Clearing and Staining Protocols for Whole-Body Cell Profiling

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