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High-fidelity detection and sorting of nanoscale vesicles in viral disease and cancer.
Morales-Kastresana, Aizea; Musich, Thomas A; Welsh, Joshua A; Telford, William; Demberg, Thorsten; Wood, James C S; Bigos, Marty; Ross, Carley D; Kachynski, Aliaksander; Dean, Alan; Felton, Edward J; Van Dyke, Jonathan; Tigges, John; Toxavidis, Vasilis; Parks, David R; Overton, W Roy; Kesarwala, Aparna H; Freeman, Gordon J; Rosner, Ariel; Perfetto, Stephen P; Pasquet, Lise; Terabe, Masaki; McKinnon, Katherine; Kapoor, Veena; Trepel, Jane B; Puri, Anu; Kobayashi, Hisataka; Yung, Bryant; Chen, Xiaoyuan; Guion, Peter; Choyke, Peter; Knox, Susan J; Ghiran, Ionita; Robert-Guroff, Marjorie; Berzofsky, Jay A; Jones, Jennifer C.
Affiliation
  • Morales-Kastresana A; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Musich TA; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Welsh JA; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Telford W; Laboratory of Pathology, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Demberg T; Experimental Immunology and Transplantation Branch, National Cancer Institute, NIH, Bethesda, MD, USA.
  • Wood JCS; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Bigos M; Wake Forest School of Medicine Flow Cytometry Core, Winston Salem, NC, USA.
  • Ross CD; Stanford University School of Medicine, Stanford, CA, USA.
  • Kachynski A; Beckman Coulter, Fort Collins, CO, USA.
  • Dean A; Beckman Coulter, Fort Collins, CO, USA.
  • Felton EJ; Beckman Coulter, Fort Collins, CO, USA.
  • Van Dyke J; Beth Israel Deaconess Medical Center, Boston, MA, USA.
  • Tigges J; University of California, Davis, Sacramento, CA, USA.
  • Toxavidis V; Beth Israel Deaconess Medical Center, Boston, MA, USA.
  • Parks DR; Beth Israel Deaconess Medical Center, Boston, MA, USA.
  • Overton WR; Stanford University School of Medicine, Stanford, CA, USA.
  • Kesarwala AH; QuantaCyte Corporation, NJ, USA.
  • Freeman GJ; Radiation Oncology Branch, National Cancer Institute, NIH, Bethesda, MD, USA.
  • Rosner A; Dana-Farber Cancer Institute, Boston, MA, USA.
  • Perfetto SP; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Pasquet L; Vaccine Research Center, National Institute of Allergy and Infectious Disease, NIH, Bethesda, MD, USA.
  • Terabe M; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • McKinnon K; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Kapoor V; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Trepel JB; Experimental Immunology and Transplantation Branch, National Cancer Institute, NIH, Bethesda, MD, USA.
  • Puri A; Developmental Therapeutics Branch, National Cancer Institute, NIH, Bethesda, MD, USA.
  • Kobayashi H; Basic Research Lab, National Cancer Institute, NIH, Frederick, MD, USA.
  • Yung B; Molecular Imaging Program, National Cancer Institute, NIH, Bethesda, MD, USA.
  • Chen X; Theranostic Nanomedicine Section, National Institute of Biomedical Imaging and Bioengineering, NIH, Bethesda, MD, USA.
  • Guion P; Theranostic Nanomedicine Section, National Institute of Biomedical Imaging and Bioengineering, NIH, Bethesda, MD, USA.
  • Choyke P; Radiation Oncology Branch, National Cancer Institute, NIH, Bethesda, MD, USA.
  • Knox SJ; Molecular Imaging Program, National Cancer Institute, NIH, Bethesda, MD, USA.
  • Ghiran I; Stanford University School of Medicine, Stanford, CA, USA.
  • Robert-Guroff M; Beth Israel Deaconess Medical Center, Boston, MA, USA.
  • Berzofsky JA; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
  • Jones JC; Vaccine Branch, National Cancer Institute, National Institutes of Health (NIH), Bethesda, MD, USA.
J Extracell Vesicles ; 8(1): 1597603, 2019.
Article in En | MEDLINE | ID: mdl-31258878
ABSTRACT
Biological nanoparticles, including viruses and extracellular vesicles (EVs), are of interest to many fields of medicine as biomarkers and mediators of or treatments for disease. However, exosomes and small viruses fall below the detection limits of conventional flow cytometers due to the overlap of particle-associated scattered light signals with the detection of background instrument noise from diffusely scattered light. To identify, sort, and study distinct subsets of EVs and other nanoparticles, as individual particles, we developed nanoscale Fluorescence Analysis and Cytometric Sorting (nanoFACS) methods to maximise information and material that can be obtained with high speed, high resolution flow cytometers. This nanoFACS method requires analysis of the instrument background noise (herein defined as the "reference noise"). With these methods, we demonstrate detection of tumour cell-derived EVs with specific tumour antigens using both fluorescence and scattered light parameters. We further validated the performance of nanoFACS by sorting two distinct HIV strains to >95% purity and confirmed the viability (infectivity) and molecular specificity (specific cell tropism) of biological nanomaterials sorted with nanoFACS. This nanoFACS method provides a unique way to analyse and sort functional EV- and viral-subsets with preservation of vesicular structure, surface protein specificity and RNA cargo activity.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Type of study: Diagnostic_studies Language: En Journal: J Extracell Vesicles Year: 2019 Document type: Article Affiliation country: Estados Unidos
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Full text: 1 Collection: 01-internacional Database: MEDLINE Type of study: Diagnostic_studies Language: En Journal: J Extracell Vesicles Year: 2019 Document type: Article Affiliation country: Estados Unidos