High Sensitivity Detection of Extracellular Vesicles PS-captured by Conventional Flow Cytometry
PRESENTER:
Dr. Naomi Tsurutani | Sales Manager at FUJIFILM Wako Laboratory Chemicals
PRESENTER'S BIO:
Naomi Tsurutani received her Ph.D. in Virology/Immunology from Tokyo Medical and Dental University in Tokyo, Japan. Before joining FUJIFILM Wako, she worked at Emory University, UMass Medical School, and UConn Health as an immunology researcher. At FUJIFILM Wako, she helps researchers to better perform and communicate their research by using FUJIFILM Wako products.
ABSTRACT:
Extracellular vesicles (EVs) such as exosomes and microvesicles serve as messengers of the intercellular networks, allowing exchange of cellular components between cells. EVs carry lipids, proteins, and nucleic acids derived from their producing cells, and have potential as biomarkers specific to cell types and cellular states. Flow cytometry is a technique that is generally used for cell marker analysis and sorting; however, analysis of small-sized EVs (50 nm to 150 nm) can be extremely challenging and frustrating. Here, we have developed a flow cytometric analysis method for Exosome markers using Tim4 immobilized magnetic beads, utilizing the property of PS binding to Exosome through phosphatidylserine (PS) (PS affinity*). In this method, first, Exosome is isolated from the sample using Tim4 immobilized magnetic beads. Next, a fluorescence-labeled Exosome marker antibody is bound to the Exosome bound to the magnetic beads via Tim4. Finally, the complex of magnetic beads, Exosome, and fluorescent-labeled antibody is analyzed by flow cytometry to detect the Exosome marker. The feature of this method is that the whole process from isolation to the detection of Exosome takes only about 2.5 hours, and highly reproducible results can be obtained by a simple experimental operation. Furthermore, it can be mentioned that the Exosome marker can be detected with high sensitivity as compared with the case of using magnetic beads on which the Exosome marker antibody is immobilized.