Imaging extracellular vesicles: current and emerging methods

Chuo, Steven Ting-Yu; Chien, Jasper Che-Yung; Lai, Charles P.

doi:10.1186/s12929-018-0494-5

Cited by 264 publications

(239 citation statements)

References 104 publications

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“…The analysis of these probes is, however, limited by the potential transfer of these dyes to other extracellular components, and their aggregation, which leads to micelle formation and unspecific labeling of acceptor cells [15,16]. Although the expression of stable fluorescent-proteins such as GFP or RFP seems to be an alternative to overcome these problems, their limited fluorescence intensity and light spectrum usually leads to low penetration and difficulties in their analysis in vivo [17]. Furthermore, fluorescence permits relative but not quantitative measurements, and is greatly affected by numerous external factors, including oxidation, scattering and bleaching [18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Gold nanoparticle based double-labeling of melanoma extracellular vesicles to determine the specificity of uptake by cells and preferential accumulation in small metastatic lung tumors

Lara¹,

Palma-Florez²,

Salas-Huenuleo³

et al. 2020

J Nanobiotechnol

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Background: Extracellular vesicles (EVs) have shown great potential for targeted therapy, as they have a natural ability to pass through biological barriers and, depending on their origin, can preferentially accumulate at defined sites, including tumors. Analyzing the potential of EVs to target specific cells remains challenging, considering the unspecific binding of lipophilic tracers to other proteins, the limitations of fluorescence for deep tissue imaging and the effect of external labeling strategies on their natural tropism. In this work, we determined the cell-type specific tropism of B16F10-EVs towards cancer cell and metastatic tumors by using fluorescence analysis and quantitative gold labeling measurements. Surface functionalization of plasmonic gold nanoparticles was used to promote indirect labeling of EVs without affecting size distribution, polydispersity, surface charge, protein markers, cell uptake or in vivo biodistribution. Double-labeled EVs with gold and fluorescent dyes were injected into animals developing metastatic lung nodules and analyzed by fluorescence/computer tomography imaging, quantitative neutron activation analysis and gold-enhanced optical microscopy. Results: We determined that B16F10 cells preferentially take up their own EVs, when compared with colon adenocarcinoma, macrophage and kidney cell-derived EVs. In addition, we were able to detect the preferential accumulation of B16F10 EVs in small metastatic tumors located in lungs when compared with the rest of the organs, as well as their precise distribution between tumor vessels, alveolus and tumor nodules by histological analysis. Finally, we observed that tumor EVs can be used as effective vectors to increase gold nanoparticle delivery towards metastatic nodules.

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Section: Introductionmentioning

confidence: 99%

Gold nanoparticle based double-labeling of melanoma extracellular vesicles to determine the specificity of uptake by cells and preferential accumulation in small metastatic lung tumors

Lara¹,

Palma-Florez²,

Salas-Huenuleo³

et al. 2020

J Nanobiotechnol

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“…[2][3][4][5] Recently, there has also been an increased interest in using lipophilic dyes to efficiently and reproducibly label both synthetic and biological nanoscale lipid vesicles such as liposomes, 6 cell-secreted extracellular vesicles 7 and viruses. [6][7][8][9][10][11][12][13][14] The photophysical properties of lipophilic membrane dyes are often greatly affected by the chemical environment and the surrounding lipids. [15][16][17] Moreover, the exact lipid composition of the membrane, including its charge and fluidity, can strongly influence the insertion efficiency of the dye and hence its effective concentration in a particular lipid vesicle.…”

Section: Introductionmentioning

confidence: 99%

Lipid vesicle composition influences the incorporation and fluorescence properties of the lipophilic sulphonated carbocyanine dye SP-DiO

Lubart

Hannestad

Pace

et al. 2020

Phys. Chem. Chem. Phys.

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“…We anticipate this work will have application in areas where capture and analysis of individual nanospecimens is of importance. For example, the optical examination of extracellular vesicles (exosomes, microvesicles, and apoptotic bodies), [ 85 ] may be facilitated by confining them in discrete points within the 2D plane of a nanocavity array. Furthermore, our technique may be used to filter particles with diameters less than the cavity diameter, allowing deterministic size‐based separation of, e.g., extracellular vesicles which have different physiological functions at different sizes.…”

Section: Resultsmentioning

confidence: 99%

Massively Multiplexed Submicron Particle Patterning in Acoustically Driven Oscillating Nanocavities

Tayebi

O’Rorke

Wong

et al. 2020

Small

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high-throughput particle manipulation for preparation, enrichment, and quality control. For example, the size distribution of synthesized nanoparticle catalysts determines their catalysis efficiency. [7,8] In biomedical applications, exosomes, which are submicron extracellular vesicles [9][10][11] containing DNA, microRNAs, and proteins, are a potential source of diagnostic biomarkers. [12][13][14][15] Suitable nanoparticle manipulation tools would enable sizebased sorting/filtering, or manipulation toward downstream activities such as multiplexed diagnostics, nanoelectronic/ robotic constructions, [16,17] and nanoparticle-based drug delivery systems. [18] Conventional particle separation techniques [19] are based on differential density and size, e.g., ultracentrifugation, [20] ultrafiltration, [21] and size exclusion chromatography. [22] Although impressive progress has been made in the last decade, many nanoparticle manipulation tools suffer from high cost, low throughput, and specimen damage. [23][24][25] Scalable nano-and submicron particle patterning has thus traditionally relied on self-assembly methods [26] or oligomer templating [27] however, these are largely unable to produce deterministic singlenanoparticle positioning, they are irreversible, and they require specific reagents or lengthy processing. [28][29][30] Moreover, drug delivery systems which incorporate efficient nanoparticle concentration are creating new avenues for cancer treatment. [31,32] There is, therefore, a pressing need to develop a fast, precise, and scalable method for nano-and submicron scale manipulation.Microfluidic devices have emerged as a viable platform for particle manipulation using magnetic, [33] optoelectronic, [34] plasmonic, [35] electrokinetic, [36] and hydrodynamic [37] forces. Applying these principles at the nanoscale instead of the microscale, however, can result in far smaller force magnitudes, poor separation efficiency, and channel clogging. [38] Furthermore, most of these manipulation methods are constrained by their dependence on particle polarizability, the need for low conductivity medium (which is often not biocompatible), and heating of the surrounding fluid. Optical tweezers [39,40] provide the highest degree of spatial resolution but they are usually applied in static fluids, owing to the relatively small forces that can be generated, [41] and only small numbers of nanoparticles can be manipulated at any one time (those within the Nanoacoustic fields are a promising method for particle actuation at the nanoscale, though THz frequencies are typically required to create nanoscale wavelengths. In this work, the generation of robust nanoscale force gradients is demonstrated using MHz driving frequencies via acoustic-structure interactions. A structured elastic layer at the interface between a microfluidic channel and a traveling surface acoustic wave (SAW) device results in submicron acoustic traps, each of which can trap individual submicron particles. The acoustically driven deformation of nanocavities gi...

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Imaging extracellular vesicles: current and emerging methods

Cited by 264 publications

References 104 publications

Gold nanoparticle based double-labeling of melanoma extracellular vesicles to determine the specificity of uptake by cells and preferential accumulation in small metastatic lung tumors

Gold nanoparticle based double-labeling of melanoma extracellular vesicles to determine the specificity of uptake by cells and preferential accumulation in small metastatic lung tumors

Lipid vesicle composition influences the incorporation and fluorescence properties of the lipophilic sulphonated carbocyanine dye SP-DiO

Massively Multiplexed Submicron Particle Patterning in Acoustically Driven Oscillating Nanocavities

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