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 language | English (US) |
|---|---|
| Pages (from-to) | 5357-5363 |
| Number of pages | 7 |
| Journal | Analytical Chemistry |
| Volume | 89 |
| Issue number | 10 |
| DOIs | |
| State | Published - May 16 2017 |
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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 journal › Article
}
TY - JOUR
T1 - Multispectral Optical Tweezers for Biochemical Fingerprinting of CD9-Positive Exosome Subpopulations
AU - Carney, Randy
AU - Hazari, Sidhartha
AU - Colquhoun, Macalistair
AU - Tran, Di
AU - Hwang, Billanna
AU - Mulligan, Michael S.
AU - Bryers, James D.
AU - Girda, Eugenia
AU - Leiserowitz, Gary S
AU - Smith, Zachary J.
AU - Lam, Kit
PY - 2017/5/16
Y1 - 2017/5/16
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85020639137&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85020639137&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.7b00017
DO - 10.1021/acs.analchem.7b00017
M3 - Article
C2 - 28345878
AN - SCOPUS:85020639137
VL - 89
SP - 5357
EP - 5363
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 10
ER -