Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations

Randy Carney, Sidhartha Hazari, Macalistair Colquhoun, Di Tran, Billanna Hwang, Michael S. Mulligan, James D. Bryers, Eugenia Girda, Gary S Leiserowitz, Zachary J. Smith, Kit Lam

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Extracellular vesicles (EVs), including exosomes, are circulating nanoscale particles heavily implicated in cell signaling and can be isolated in vast numbers from human biofluids. Study of their molecular profiling and materials properties is currently underway for purposes of describing a variety of biological functions and diseases. However, the large, and as yet largely unquantified, variety of EV subpopulations differing in composition, size, and likely function necessitates characterization schemes capable of measuring single vesicles. Here we describe the first application of multispectral optical tweezers (MS-OTs) to single vesicles for molecular fingerprinting of EV subpopulations. This versatile imaging platform allows for sensitive measurement of Raman chemical composition (e.g., variation in protein, lipid, cholesterol, nucleic acids), coupled with discrimination by fluorescence markers. For exosomes isolated by ultracentrifugation, we use MS-OTs to interrogate the CD9-positive subpopulations via antibody fluorescence labeling and Raman spectra measurement. We report that the CD9-positive exosome subset exhibits reduced component concentration per vesicle and reduced chemical heterogeneity compared to the total purified EV population. We observed that specific vesicle subpopulations are present across exosomes isolated from cell culture supernatant of several clonal varieties of mesenchymal stromal cells and also from plasma and ascites isolated from human ovarian cancer patients.

Original languageEnglish (US)
Pages (from-to)5357-5363
Number of pages7
JournalAnalytical Chemistry
Volume89
Issue number10
DOIs
StatePublished - May 16 2017

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Optical tweezers
Fluorescence
Cell signaling
Chemical analysis
Cell culture
Labeling
Nucleic Acids
Raman scattering
Materials properties
Cholesterol
Lipids
Plasmas
Imaging techniques
Antibodies
Proteins
Ultracentrifugation

ASJC Scopus subject areas

  • Analytical Chemistry

Cite this

Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations. / Carney, Randy; Hazari, Sidhartha; Colquhoun, Macalistair; Tran, Di; Hwang, Billanna; Mulligan, Michael S.; Bryers, James D.; Girda, Eugenia; Leiserowitz, Gary S; Smith, Zachary J.; Lam, Kit.

In: Analytical Chemistry, Vol. 89, No. 10, 16.05.2017, p. 5357-5363.

Research output: Contribution to journalArticle

Carney, R, Hazari, S, Colquhoun, M, Tran, D, Hwang, B, Mulligan, MS, Bryers, JD, Girda, E, Leiserowitz, GS, Smith, ZJ & Lam, K 2017, 'Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations', Analytical Chemistry, vol. 89, no. 10, pp. 5357-5363. https://doi.org/10.1021/acs.analchem.7b00017
Carney, Randy ; Hazari, Sidhartha ; Colquhoun, Macalistair ; Tran, Di ; Hwang, Billanna ; Mulligan, Michael S. ; Bryers, James D. ; Girda, Eugenia ; Leiserowitz, Gary S ; Smith, Zachary J. ; Lam, Kit. / Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations. In: Analytical Chemistry. 2017 ; Vol. 89, No. 10. pp. 5357-5363.
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