Academic Journal
The fluorochrome-to-protein ratio is crucial for the flow cytometric detection of tissue factor on extracellular vesicles
| العنوان: | The fluorochrome-to-protein ratio is crucial for the flow cytometric detection of tissue factor on extracellular vesicles |
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| المؤلفون: | René Weiss, Marwa Mostageer, Tanja Eichhorn, Silke Huber, Dominik Egger, Andreas Spittler, Carla Tripisciano, Cornelia Kasper, Viktoria Weber |
| المصدر: | Scientific Reports, Vol 14, Iss 1, Pp 1-10 (2024) |
| بيانات النشر: | Nature Portfolio, 2024. |
| سنة النشر: | 2024 |
| المجموعة: | LCC:Medicine LCC:Science |
| مصطلحات موضوعية: | Extracellular vesicles, Flow cytometry, Fluorochrome-to-protein ratio, Tissue factor, Medicine, Science |
| الوصف: | Abstract Extracellular vesicles (EVs) have crucial roles in hemostasis and coagulation. They sustain coagulation by exposing phosphatidylserine and initiate clotting by surface expression of tissue factor (TF) under inflammatory conditions. As their relevance as biomarkers of coagulopathy is increasingly recognized, there is a need for the sensitive and reliable detection of TF+ EVs, but their flow cytometric analysis is challenging and has yielded controversial findings for TF expression on EVs in the vascular system. We investigated the effect of different fluorochrome-to-protein (F/P) ratios of anti-TF-fluorochrome conjugates on the flow cytometric detection of TF+ EVs from activated monocytes, mesenchymal stem cells (MSCs), and in COVID-19 plasma. Using a FITC-labeled anti-TF antibody (clone VD8), we show that the percentage of TF+ EVs declined with decreasing F/P ratios. TF was detected on 7.6%, 5.4%, and 1.1% of all EVs derived from activated monocytes at F/P ratios of 7.7:1, 6.6:1, and 5.2:1. A similar decline was observed for EVs from MSCs and for EVs in plasma, whereas the detection of TF on cells remained unaffected by different F/P ratios. We provide clear evidence that next to the antibody clone, the F/P ratio affects the flow cytometric detection of TF+ EVs and should be carefully controlled. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 2045-2322 |
| Relation: | https://doaj.org/toc/2045-2322 |
| DOI: | 10.1038/s41598-024-56841-5 |
| URL الوصول: | https://doaj.org/article/1606774928954ae28a76785d00402c14 |
| رقم الانضمام: | edsdoj.1606774928954ae28a76785d00402c14 |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 20452322 |
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| DOI: | 10.1038/s41598-024-56841-5 |