A nano flow cytometer for single lipid vesicle analysis
Artikel i vetenskaplig tidskrift, 2017

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.

UMR168

membrane curvature

Paris

p8477

Biochemistry & Molecular Biology

RS

France.

Chemistry

emerging role

immune-responses

nanoparticle tracking analysis

extracellular vesicles

individual nanoscale liposomes

Other Topics

cell-penetrating peptides

Science & Technology -

amphipathic helices

live cells

fluorescence

v109

ates of america

Physicochim Curie

Författare

R. Friedrich

Chalmers University of Technology

Stephan Block

Chalmers, Fysik, Biologisk fysik

Mohammadreza Alizadehheidari

Chalmers, Biologi och bioteknik, Kemisk biologi

Susanne Heider

Chalmers, Biologi och bioteknik, Kemisk biologi

Joachim Fritzsche

Chalmers, Fysik, Kemisk fysik

Elin Esbjörner Winters

Chalmers, Biologi och bioteknik, Kemisk biologi

Fredrik Westerlund

Chalmers, Biologi och bioteknik, Kemisk biologi

Marta Bally

Chalmers, Fysik, Biologisk fysik

Lab on a Chip - Miniaturisation for Chemistry and Biology

1473-0197 (ISSN) 1473-0189 (eISSN)

Vol. 17 830-841

Ämneskategorier

Biofysik

DOI

10.1039/c6lc01302c

PubMed

28128381