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Comparison and opti...
Comparison and optimization of nanoscale extracellular vesicle imaging by scanning electron microscopy for accurate size-based profiling and morphological analysis
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- Cavallaro, Sara (författare)
- KTH,Fotonik,Department of Applied Physics;School of Engineering Sciences;KTH Royal Institute of Technology;10691 Stockholm;Sweden,KTH Royal Inst Technol, Sch Engn Sci, Dept Appl Phys, S-10691 Stockholm, Sweden
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- Hååg, Petra (författare)
- Karolinska Institutet,Karolinska Inst, Dept Oncol Pathol, S-17164 Solna, Sweden
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- Viktorsson, Kristina (författare)
- Karolinska Institutet,Karolinska Inst, Dept Oncol Pathol, S-17164 Solna, Sweden
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- Krozer, Anatol (författare)
- RISE,Smart hårdvara,Department of Smart Hardware;Division of Digital Systems;Research Institutes of Sweden AB;40014 Gothenburg;Sweden,Res Inst Sweden AB, Div Digital Syst, Dept Smart Hardware, S-40014 Gothenburg, Sweden
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- Fogel, Kristina (författare)
- RISE,Smart hårdvara,Department of Smart Hardware;Division of Digital Systems;Research Institutes of Sweden AB;40014 Gothenburg;Sweden,Res Inst Sweden AB, Div Digital Syst, Dept Smart Hardware, S-40014 Gothenburg, Sweden
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- Lewensohn, Rolf (författare)
- Karolinska Institutet,Karolinska Inst, Dept Oncol Pathol, S-17164 Solna, Sweden; Karolinska Univ Hosp, Thorac Oncol Ctr, Theme Canc, Med Unit Head & Neck Lung & Skin Tumors, S-17164 Solna, Sweden
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- Linnros, Jan, 1953- (författare)
- KTH,Fotonik,KTH Royal Inst Technol, Sch Engn Sci, Dept Appl Phys, S-10691 Stockholm, Sweden
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- Dev, Apurba (författare)
- Uppsala universitet,KTH,Fotonik,Fasta tillståndets elektronik,KTH Royal Inst Technol, Sch Engn Sci, Dept Appl Phys, S-10691 Stockholm, Sweden
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(creator_code:org_t)
- 2021
- 2021
- Engelska.
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Ingår i: Nanoscale Advances. - : Royal Society of Chemistry. - 2516-0230. ; 3:11, s. 3053-3063
- Relaterad länk:
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https://doi.org/10.1...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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http://kipublication...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- Nanosized extracellular vesicles (EVs) have been found to play a key role in intercellular communication, offering opportunities for both disease diagnostics and therapeutics. However, lying below the diffraction limit and also being highly heterogeneous in their size, morphology and abundance, these vesicles pose significant challenges for physical characterization. Here, we present a direct visual approach for their accurate morphological and size-based profiling by using scanning electron microscopy (SEM). To achieve that, we methodically examined various process steps and developed a protocol to improve the throughput, conformity and image quality while preserving the shape of EVs. The study was performed with small EVs (sEVs) isolated from a non-small-cell lung cancer (NSCLC) cell line as well as from human serum, and the results were compared with those obtained from nanoparticle tracking analysis (NTA). While the comparison of the sEV size distributions showed good agreement between the two methods for large sEVs (diameter > 70 nm), the microscopy based approach showed a better capacity for analyses of smaller vesicles, with higher sEV counts compared to NTA. In addition, we demonstrated the possibility of identifying non-EV particles based on size and morphological features. The study also showed process steps that can generate artifacts bearing resemblance with sEVs. The results therefore present a simple way to use a widely available microscopy tool for accurate and high throughput physical characterization of EVs.
Ämnesord
- NATURVETENSKAP -- Biologi -- Cellbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Cell Biology (hsv//eng)
- TEKNIK OCH TEKNOLOGIER -- Nanoteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Nano-technology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Cell- och molekylärbiologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Cell and Molecular Biology (hsv//eng)
Nyckelord
- Cell culture
- Diagnosis
- Diffraction
- Image enhancement
- Scanning electron microscopy
- Diffraction limits
- Disease diagnostics
- Intercellular communications
- Morphological analysis
- Morphological features
- Nanoparticle tracking analysis
- Non small cell lung cancer
- Physical characterization
- Particle size analysis
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- art (ämneskategori)
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