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A nano flow cytomet...
A nano flow cytometer for single lipid vesicle analysis
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- Friedrich, R. (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Block, Stephan, 1978 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Alizadehheidari, Mohammadreza, 1987 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Heider, Susanne, 1984 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Fritzsche, Joachim, 1977 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Esbjörner Winters, Elin, 1978 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Westerlund, Fredrik, 1978 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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- Bally, Marta, 1981 (author)
- Chalmers tekniska högskola,Chalmers University of Technology
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(creator_code:org_t)
- 2017
- 2017
- English.
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In: Lab on a Chip - Miniaturisation for Chemistry and Biology. - : Royal Society of Chemistry (RSC). - 1473-0189 .- 1473-0197. ; 17:5, s. 830-841
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Abstract
Subject headings
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- We present a nanofluidic device for fluorescence-based detection and characterization of small lipid vesicles on a single particle basis. The device works like a nano flow cytometer where individual vesicles are visualized by fluorescence microscopy while passing through parallel nanochannels in a pressure-driven flow. An experiment requires less than 20 mu l sample volume to quantify both the vesicle content and the fluorescence signals emitted by individual vesicles. We show that the device can be used to accurately count the number of fluorescent synthetic lipid vesicles down to a vesicle concentration of 170 fM. We also show that the size-distribution of the vesicles can be resolved from their fluorescence intensity distribution after calibration. We demonstrate the applicability of the assay in two different examples. In the first, we use the nanofluidic device to determine the particle concentration in a sample containing cell-derived extracellular vesicles labelled with a lipophilic dye. In the second, we demonstrate that dual-color detection can be used to probe peptide binding to synthetic lipid vesicles; we identify a positive membrane-curvature sensing behavior of an arginine enriched version of the Antennapedia homeodomain peptide penetratin. Altogether, these results illustrate the potential of this nanofluidic-based methodology for characterization and quantification of small biological vesicles and their interactors without ensemble averaging. The device is therefore likely to find use as a quantitative analytical tool in a variety of fields ranging from diagnostics to fundamental biology research. Moreover, our results have potential to facilitate further development of automated lab-on-a-chip devices for vesicle analysis.
Subject headings
- NATURVETENSKAP -- Biologi -- Biofysik (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biophysics (hsv//eng)
Keyword
- UMR168
- ates of america
- France.
- v109
- individual nanoscale liposomes
- cell-penetrating peptides
- fluorescence
- RS
- membrane curvature
- Physicochim Curie
- nanoparticle tracking analysis
- immune-responses
- Biochemistry & Molecular Biology
- p8477
- live cells
- Paris
- emerging role
- Science & Technology -
- amphipathic helices
- extracellular vesicles
- Other Topics
- Chemistry
Publication and Content Type
- art (subject category)
- ref (subject category)
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- By the author/editor
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Friedrich, R.
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Block, Stephan, ...
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Alizadehheidari, ...
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Heider, Susanne, ...
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Fritzsche, Joach ...
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Esbjörner Winter ...
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Westerlund, Fred ...
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Bally, Marta, 19 ...
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- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
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and Biophysics
- Articles in the publication
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Lab on a Chip - ...
- By the university
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Chalmers University of Technology