Quantum Dot Labeling and Visualization of Extracellular Vesicles

Zhang, Mengying; Vojtech, Lucia; Ye, Ziming; Hladik, Florian; Nance, Elizabeth

doi:10.1021/acsanm.0c01553

Cited by 41 publications

(47 citation statements)

References 70 publications

(130 reference statements)

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“…Cadmium selenide (CdSe) based quantum dots modified with polyethylene glycol and chemically linked to interleukin-13 (IL13QD) were prepared with the aim to identify the binding capacity of IL13QD to EVs from the CSF of the brain tumour patients isolated by differential centrifugation followed by UC [111]. Quite intriguing is also a recent study on EVs of seminal origin, in which quantum dots were covalently bonded to the primary amines on the surface of EVs via click chemistry [112]. This enabled their identification in TEM as well as a stable fluorescent labelling and live tracking.…”

Section: Application Of Em For Characterisation Of Evs From Human Biofluidsmentioning

confidence: 99%

Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids

et al. 2021

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Extracellular vesicles (EVs) are nanometric membranous structures secreted from almost every cell and present in biofluids. Because EV composition reflects the state of its parental tissue, EVs possess an enormous diagnostic/prognostic potential to reveal pathophysiological conditions. However, a prerequisite for such usage of EVs is their detailed characterisation, including visualisation which is mainly achieved by atomic force microscopy (AFM) and electron microscopy (EM). Here we summarise the EV preparation protocols for AFM and EM bringing out the main challenges in the imaging of EVs, both in their natural environment as biofluid constituents and in a saline solution after EV isolation. In addition, we discuss approaches for EV imaging and identify the potential benefits and disadvantages when different AFM and EM methods are applied, including numerous factors that influence the morphological characterisation, standardisation, or formation of artefacts. We also demonstrate the effects of some of these factors by using cerebrospinal fluid as an example of human biofluid with a simpler composition. Here presented comparison of approaches to EV imaging should help to estimate the current state in morphology research of EVs from human biofluids and to identify the most efficient pathways towards the standardisation of sample preparation and microscopy modes.

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Section: Application Of Em For Characterisation Of Evs From Human Biofluidsmentioning

confidence: 99%

Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids

et al. 2021

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“…Direct surface conjugation was mainly achieved by attaching chemical linkers to exosome surfaces via covalent binding to surface proteins [ 25 ] or hydrophobic tail insertion of molecules with click chemistry to lipid bilayer of exosomes [ 26 ]. These functional linkers subsequently reacted with the functional surfaces of nanoparticles, leading to a “raspberry” nanoconstruct where the exosomes were decorated with inorganic nanoparticles.…”

Section: Preparation Of Nanoparticle-loaded Exosomesmentioning

confidence: 99%

“…For example, a hydrazine–aldehyde-based linking strategy based on 4-formylbenzoate (4FB) to 6-hydrazinonicotinate acetone hydrazone (HyNic) click chemistry was explored to label exosomes with quantum dots (QDs) [ 25 ]. Specifically, commercially available 10–20 nm QDs functionalized with amine-terminated polyethylene glycol (QD-PEG-NH 2 ) were modified with Sulfo-S-4FB, while exosomes isolated from body fluids were labeled with Sulfo-S-HyNic through the interactions between amine-reactive NHS-esters and exosome surface proteins.…”

Section: Preparation Of Nanoparticle-loaded Exosomesmentioning

confidence: 99%

“…Subsequently, 40 nm Au nanoparticles functionalized with thiol groups were effectively attached to the surfaces of MCF-7 cell derived exosomes ( Figure 2 b). The surface conjugation strategies using chemical linkers to attach inorganic nanoparticles to exosome surfaces must be carefully selected [ 25 ] to minimize the disruption of exosome structural integrity. In addition, the water solubility of the linker is critical, because the use of organic solvents would cause potential damage to exosomes.…”

Section: Preparation Of Nanoparticle-loaded Exosomesmentioning

confidence: 99%

“… Examples of inorganic nanoparticle (NP)-loaded exosomes post exosome formation with schematic drawings and TEM images: ( a ) QD-loaded exosomes through chemical linker conjugation, [ 25 ] ( b ) Au NP-loaded exosomes through lipid infused linkers, [ 26 ] ( c ) Au NP-loaded exosomes through extrusion, [ 28 ] and ( d ) V 2 C DD-loaded exosomes via electroporation [ 30 ]. …”

Section: Figurementioning

confidence: 99%

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Inorganic Nanoparticle-Loaded Exosomes for Biomedical Applications

2021

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Exosomes are intrinsic cell-derived membrane vesicles in the size range of 40–100 nm, serving as great biomimetic nanocarriers for biomedical applications. These nanocarriers are known to bypass biological barriers, such as the blood–brain barrier, with great potential in treating brain diseases. Exosomes are also shown to be closely associated with cancer metastasis, making them great candidates for tumor targeting. However, the clinical translation of exosomes are facing certain critical challenges, such as reproducible production and in vivo tracking of their localization, distribution, and ultimate fate. Recently, inorganic nanoparticle-loaded exosomes have been shown great benefits in addressing these issues. In this review article, we will discuss the preparation methods of inorganic nanoparticle-loaded exosomes, and their applications in bioimaging and therapy. In addition, we will briefly discuss their potentials in exosome purification.

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DNA Nanomaterial‐Empowered Surface Engineering of Extracellular Vesicles

Yao,

He,

Wei

et al. 2023

Advanced Materials

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Extracellular vesicles (EVs) are cell‐secreted biological nanoparticles that are critical mediators of intercellular communication. They contain diverse bioactive components, which are promising diagnostic biomarkers and therapeutic agents. Their nanosized membrane‐bound structures and innate ability to transport functional cargo across major biological barriers make them promising candidates as drug delivery vehicles. However, the complex biology and heterogeneity of EVs pose significant challenges for their controlled and actionable applications in diagnostics and therapeutics. Recently, DNA molecules with high biocompatibility emerge as excellent functional blocks for surface engineering of EVs. The robust Watson–Crick base pairing of DNA molecules and the resulting programmable DNA nanomaterials provide the EV surface with precise structural customization and adjustable physical and chemical properties, creating unprecedented opportunities for EV biomedical applications. This review focuses on the recent advances in the utilization of programmable DNA to engineer EV surfaces. The biology, function, and biomedical applications of EVs are summarized and the state‐of‐the‐art achievements in EV isolation, analysis, and delivery based on DNA nanomaterials are introduced. Finally, the challenges and new frontiers in EV engineering are discussed.

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Quantum Dot Labeling and Visualization of Extracellular Vesicles

Cited by 41 publications

References 70 publications

Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids

Perspectives of Microscopy Methods for Morphology Characterisation of Extracellular Vesicles from Human Biofluids

Inorganic Nanoparticle-Loaded Exosomes for Biomedical Applications

DNA Nanomaterial‐Empowered Surface Engineering of Extracellular Vesicles

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