Sizing and phenotyping of cellular vesicles using Nanoparticle Tracking Analysis

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• 作者：Rebecca A.  ; Dragovic MSc^a ;   ; ¹ ; Christopher  ; Gardiner PhD^a ;   ; ¹ ; Alexandra S.  ; Brooks MD^a ; Dionne S.  ; Tannetta PhD^a ; David J.P.  ; Ferguson PhD^b ; Patrick  ; Hole PhD^c ; Bob  ; Carr PhD^c ; Christopher W.G.  ; Redman MD ; FRCP^a ; Adrian L.  ; Harris MD ; PhD ; FRCP^d ; Peter J.  ; Dobson PhD ; FInstP ; CPhys^e ; Paul  ; Harrison PhD^f ; Ian L.  ; Sargent PhD^a ;   ; ^{ian.sargent@obs-gyn.ox.ac.uk}
• 关键词：Microvesicles ; Microparticles ; Exosomes ; Nanoparticle Tracking Analysis ; Flow Cytometry
• 刊名：Nanomedicine: Nanotechnology, Biology and Medicine
• 出版年：2011
• 出版时间：December 2011
• 年：2011
• 卷：7
• 期：6
• 页码：780-788
• 全文大小：897 K

文摘

Cellular microvesicles and nanovesicles (exosomes) are involved in many disease processes and have major potential as biomarkers. However, developments in this area are constrained by limitations in the technology available for their measurement. Here we report on the use of fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles. In this system vesicles are visualized by light scattering using a light microscope. A video is taken, and the NTA software tracks the brownian motion of individual vesicles and calculates their size and total concentration. Using human placental vesicles and plasma, we have demonstrated that NTA can measure cellular vesicles as small as ?0 nm and is far more sensitive than conventional flow cytometry (lower limit ?00 nm). By combining NTA with fluorescence measurement we have demonstrated that vesicles can be labeled with specific antibody-conjugated quantum dots, allowing their phenotype to be determined.

From the Clinical Editor

The authors of this study utilized fluorescence nanoparticle tracking analysis (NTA) to rapidly size and phenotype cellular vesicles, demonstrating that NTA is far more sensitive than conventional flow cytometry.