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High‐Throughput Counting and Superresolution Mapping of Tetraspanins on Exosomes Using a Single‐Molecule Sensitive Flow Technique and Transistor‐like Semiconducting Polymer Dots
Author(s) -
Jiang Yifei,
Andronico Luca A.,
Jung SeungRyoung,
Chen Haobin,
Fujimoto Bryant,
Vojtech Lucia,
Chiu Daniel T.
Publication year - 2021
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.202103282
Subject(s) - microvesicles , materials science , microfluidics , throughput , nanotechnology , transistor , exosome , polymer , chemistry , computer science , physics , telecommunications , microrna , biochemistry , composite material , quantum mechanics , voltage , wireless , gene
A method for high‐throughput counting and superresolution mapping of surface proteins on exosomes is described. The method combines a single‐molecule sensitive flow technique and an adaptive superresolution imaging method. Exosomes stained with membrane dye and dye‐conjugated antibodies were analyzed using a microfluidic platform at a flow rate of 100 exosome s −1 to determine size and protein copy number. Superresolution mapping was performed with exosomes labeled with novel transistor‐like, semiconducting polymer dots (Pdots), which exhibit spontaneous blinking with <5 nm localization error and a broad range of optical‐adjustable duty cycles. Based on the copy numbers extracted from the flow analysis, the switch‐on frequency of the Pdots were finely adjusted so that structures of hundreds of exosomes were obtained within five minutes. The high throughput and high sensitivity of this method offer clear advantages for characterization of exosomes and similar biological vesicles.