液体的可追溯折射率测量标准化细胞外囊泡流细胞计
Martine Kuiper1,2,3,4, Richard Koops1, Rienk Nieuwland2,3
1VSL, National Metrology Institute, Delft, the Netherlands.
概括
使用流式细胞计量精确测量细胞外囊泡 (EV) 度,需要精确的折射率 (RI) 值悬浮液体. 使用可追溯的RI可以提高EV大小和度量化的准确性.
科学领域:
- 生物医学工程 生物医学工程
- 分析化学 分析化学
- 生物物理学的生物物理.
背景情况:
- 细胞外囊泡 (EVs) 是体液中发现的关键生物标志物.
- 流细胞计用于量化EV度,依赖于光散射.
- 精确的尺寸和量化取决于Mie理论,这需要周围介质的精确折射率 (RI).
研究的目的:
- 用于测量用于EV分析的常见悬浮液体的可追溯折射率 (RIs).
- 评估 RI 精度对 EV 尺寸和度测量的影响.
- 为改善EV流细胞计的准确性提供建议.
主要方法:
- 可追溯测量Dulbecco的酸盐缓冲盐水 (DPBS) 和稀释的人体血的RI.
- 使用的波长在405和644nm之间,不确定性为1.4
× 10 - 5 $$ 1.4\times {10}^{-5} $$ . - 通过使用Mie理论和流细胞计,从假设与测量RI获得的EV度进行比较.
主要成果:
- 准确确定DPBS和血稀释的可追溯RI.
- 假设和测量RI之间的差异显著改变了光径门和报告的EV度.
- 在罗塞塔校准中使用目前假设的DPBS的RI导致EV度在9%到35%的差异.
结论:
- 准确地确定悬浮液体的RI对于可靠的EV量化至关重要.
- 目前在EV流细胞计校准中的RI假设导致了显著的测量错误.
- 在 Mie 理论计算中实施可追溯测量的 RI 将提高 EV 光径和度测量的准确性.
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