Protein-based fluorescent nanoparticles for super-resolution STED imaging of live cells

Li, Shang; Gao, Peng; Wang, Haixia; Popescu, Radian; Gerthsen, D.; Nienhaus, G. Ulrich

doi:10.1039/c6sc04664a

Cited by 37 publications

(31 citation statements)

References 44 publications

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“…All solutions were prepared in LoBind tubes (Eppendorf, Hamburg, Germany). Using Atto647N‐NHS‐ester (ATTO‐TEC, Siegen, Germany), dye labeling of Tf was performed as previously reported . The degree of labeling (dye/protein ratio) was determined spectrophotometrically as 34.2 ± 0.2%.…”

Section: Methodsmentioning

confidence: 99%

Formation of a Monolayer Protein Corona around Polystyrene Nanoparticles and Implications for Nanoparticle Agglomeration

Wang

Nienhaus

et al. 2019

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Nanoparticle (NP) interactions with cells and organisms are mediated by a biomolecular adsorption layer, the so‐called “protein corona.” An in‐depth understanding of the corona is a prerequisite to successful and safe application of NPs in biology and medicine. In this work, earlier in situ investigations on small NPs are extended to large polystyrene (PS) NPs of up to 100 nm diameter, using human transferrin (Tf) and human serum albumin (HSA) as model proteins. Direct NP sizing experiments reveal a reversibly bound monolayer protein shell (under saturating conditions) on hydrophilic, carboxyl‐functionalized (PS‐COOH) NPs, as was earlier observed for much smaller NPs. In contrast, protein binding on hydrophobic, sulfated (PS‐OSO3H) NPs in solvent of low ionic strength is completely irreversible; nevertheless, the thickness of the observed protein corona again corresponds to a protein monolayer. Under conditions of reduced charge repulsion (higher ionic strength), the NPs are colloidally unstable and form large clusters below a certain protein–NP stoichiometric ratio, indicating that the adsorbed proteins induce NP agglomeration. This comprehensive characterization of the persistent protein corona on PS‐OSO3H NPs by nanoparticle sizing and quantitative fluorescence microscopy/nanoscopy reveals mechanistic aspects of molecular interactions occurring during exposure of NPs to biofluids.

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Section: Methodsmentioning

confidence: 99%

Formation of a Monolayer Protein Corona around Polystyrene Nanoparticles and Implications for Nanoparticle Agglomeration

Wang

Nienhaus

et al. 2019

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“…More recently, Peuschel et al used 3D STED to quantify the internalization of 25 nm and 85 nm SNPs in A549 cells 117 and differentiate between membraneattached and internalized nanoparticles. A similar approach was also used to study the internalization and colocalization of carbon dots 118 and protein-based fluorescent nanoparticles 119 . In the latter case, nanoparticles were functionalised using 119 transferrin that was in turn labeled with a optimal STED dye (Atto647N).…”

Section: [H2] Cellular Trafficking and Localizationmentioning

confidence: 99%

Super-resolution microscopy as a powerful tool to study complex synthetic materials

et al. 2019

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“…Organic molecules have been widely used in fluorescence microscopy due to their bright emissions and excellent biocompatibility . However, organic molecules are photobleached easily under strong laser excitation, which has restricted their use for long‐term STED nanoscopy . Although photobleaching resistance can be optimized by encapsulating organic molecules within a certain matrix to form fluorescent nanoparticles (NPs), a lateral resolution of ≈30 nm based on aggregation induced emission nanoparticle‐assisted STED imaging could be obtained only with a very high‐intensity STED beam (higher than 300 mW) .…”

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confidence: 99%

Low‐Saturation‐Intensity, High‐Photostability, and High‐Resolution STED Nanoscopy Assisted by CsPbBr₃ Quantum Dots

Yan

Zhao

et al. 2018

Advanced Materials

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Stimulated emission depletion (STED) nanoscopy is one of the most promising super-resolution imaging techniques for microstructure imaging. Commercial CdSe@ZnS quantum dots are used as STED probes and ≈50 nm lateral resolution is obtained. Compared with other quantum dots, perovskite CsPbBr nanoparticles (NPs) possess higher photoluminescence quantum yield and larger absorption cross-section, making them a more effective probe for STED nanoscopy. In this study, CsPbBr NPs are used as probes for STED nanoscopy imaging. The fluorescence intensity of the CsPbBr sample is hardly weakened at all after 200 min irradiation with a 39.8 mW depletion laser, indicating excellent photobleaching resistance of the CsPbBr NPs. The saturation intensity of the CsPbBr NPs is extremely low and estimated to be only 0.4 mW (0.126 MW cm ). Finally, an ultrahigh lateral resolution of 20.6 nm is obtained for a single nanoparticle under 27.5 mW STED laser irradiation in CsPbBr -based STED nanoscopy imaging, which is a tenfold improvement compared with confocal microscopy. Because of its high fluorescence stability and ultrahigh resolution under lower depletion power, CsPbBr -assisted STED nanoscopy has great potential to investigate microstructures that require super-resolution and long-term imaging.

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Protein-based fluorescent nanoparticles for super-resolution STED imaging of live cells

Abstract: Protein-based fluorescent nanoparticles with excellent biocompatibility, good colloidal stability and photostability have been synthesized as attractive markers for STED nanoscopy in biological imaging.

Cited by 37 publications

References 44 publications

Formation of a Monolayer Protein Corona around Polystyrene Nanoparticles and Implications for Nanoparticle Agglomeration

Formation of a Monolayer Protein Corona around Polystyrene Nanoparticles and Implications for Nanoparticle Agglomeration

Super-resolution microscopy as a powerful tool to study complex synthetic materials

Low‐Saturation‐Intensity, High‐Photostability, and High‐Resolution STED Nanoscopy Assisted by CsPbBr₃ Quantum Dots

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Protein-based fluorescent nanoparticles for super-resolution STED imaging of live cells

Abstract: Protein-based fluorescent nanoparticles with excellent biocompatibility, good colloidal stability and photostability have been synthesized as attractive markers for STED nanoscopy in biological imaging.

Cited by 37 publications

References 44 publications

Formation of a Monolayer Protein Corona around Polystyrene Nanoparticles and Implications for Nanoparticle Agglomeration

Formation of a Monolayer Protein Corona around Polystyrene Nanoparticles and Implications for Nanoparticle Agglomeration

Super-resolution microscopy as a powerful tool to study complex synthetic materials

Low‐Saturation‐Intensity, High‐Photostability, and High‐Resolution STED Nanoscopy Assisted by CsPbBr3 Quantum Dots

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Low‐Saturation‐Intensity, High‐Photostability, and High‐Resolution STED Nanoscopy Assisted by CsPbBr₃ Quantum Dots