title: On-chip light sheet illumination enables diagnostic size and concentration measurements of membrane vesicles in biofluids.
abstract: Cell-derived membrane vesicles that are released in biofluids, like blood or saliva, are emerging as potential non-invasive biomarkers for diseases, such as cancer. Techniques capable of measuring the size and concentration of membrane vesicles directly in biofluids are urgently needed. Fluorescence single particle tracking microscopy has the potential of doing exactly that by labelling the membrane vesicles with a fluorescent label and analysing their Brownian motion in the biofluid. However, an unbound dye in the biofluid can cause high background intensity that strongly biases the fluorescence single particle tracking size and concentration measurements. While such background intensity can be avoided with light sheet illumination, current set-ups require specialty sample holders that are not compatible with high-throughput diagnostics. Here, a microfluidic chip with integrated light sheet illumination is reported, and accurate fluorescence single particle tracking size and concentration measurements of membrane vesicles in cell culture medium and in interstitial fluid collected from primary human breast tumours are demonstrated.
authors.affiliation: Laboratory of General Biochemistry and Physical Pharmacy, Ghent University, Belgium. Kevin.Braeckmans@UGent.be.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Deschout
authors.forename: Koen
authors.initials: K
authors.lastname: Raemdonck
authors.forename: Stephan
authors.initials: S
authors.lastname: Stremersch
authors.forename: Pietro
authors.initials: P
authors.lastname: Maoddi
authors.forename: Guillaume
authors.initials: G
authors.lastname: Mernier
authors.forename: Philippe
authors.initials: P
authors.lastname: Renaud
authors.forename: Sébastien
authors.initials: S
authors.lastname: Jiguet
authors.forename: An
authors.initials: A
authors.lastname: Hendrix
authors.forename: Marc
authors.initials: M
authors.lastname: Bracke
authors.forename: Rudy
authors.initials: R
authors.lastname: Van den Broecke
authors.forename: Magnus
authors.initials: M
authors.lastname: Röding
authors.forename: Mats
authors.initials: M
authors.lastname: Rudemo
authors.forename: Jo
authors.initials: J
authors.lastname: Demeester
authors.forename: Stefaan C
authors.initials: SC
authors.lastname: De Smedt
authors.forename: Filip
authors.initials: F
authors.lastname: Strubbe
authors.forename: Kristiaan
authors.initials: K
authors.lastname: Neyts
authors.forename: Kevin
authors.initials: K
authors.lastname: Braeckmans
mesh_terms: D016477:Artifacts; D015415:Biomarkers / Q000378:metabolism*; D015374:Biosensing Techniques / Q000379:methods*; D001943:Breast Neoplasms / Q000378:metabolism; D045744:Cell Line, Tumor; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000737:chemistry; D006801:Humans; D008027:Light; D008422:Materials Testing; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics / Q000379:methods*; D009038:Motion; D010316:Particle Size; D012542:Scattering, Radiation; D012825:Silicon / Q000737:chemistry
chemical_list: D015415:Biomarkers; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D012825:Silicon