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Fluorescence labelled antibodies are routinely used to label extracellular vesicles (EVs) to better identify EVs. Like most of the protein, antibodies tend to form aggregates to reach their equilibrium state. Antibody aggregates may have a size similar to EVs and fluoresce stronger than labelled EVs, and could potentially cause misinterpretation. To solve this problem, high-speed centrifugation or filtration method are used to remove antibody aggregates before EVs labelling has been proposed. Compared to the conventional flow cytometer, the Apogee A60 Micro-PLUS flow cytometer suits the application to detect minute size differences in small particles that differentiate labelled EVs from antibody aggregates. In the advent of Apogee A60 Micro-PLUS high-resolution flow cytometer, EVs population and antibody aggregates can be clearly distinguished and identified from the cytogram and rule out the possibility of over or under-estimation of EVs.
- Rasmussen, R.W.; Botha, J.; Prip, F.; Sanden, M.; Nielsen, M.H.; Handberg, A. Zoom in on Antibody Aggregates: A Potential Pitfall in the Search of Rare EV Populations. Biomedicines 2021, 9, 206. https://doi.org/10.3390/biomedicines9020206