Microparticle sizing by dynamic light scattering in fresh-frozen plasma
Microparticle sizing by dynamic light scattering in fresh-frozen plasma
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DOI：
10.1111/j.1423-0410.2008.01151.x
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发表时间：
2009-04-01
期刊：
影响因子：
2.7
通讯作者：
Harrison, P.
中科院分区：
文献类型：
--
作者：
Lawrie, A. S.;Albanyan, A.;Harrison, P.
我们之前已经证明，新鲜冰冻血浆(FFP)含有红细胞衍生的促凝血剂微粒(MPS)，可通过0.2微米过滤去除。鉴于目前精确测定MPS大小的方法的局限性，我们应用新的动态光散射(DLS)方法来表征这些MPS在FFP中的尺寸分布。新鲜冰冻血浆是由A型和O型献血者(n=10)在4℃下过夜保存全血后制备的。在分析当天，血浆解冻至37℃，并对过滤前和过滤后的子体等分进行研究(0.2微米过滤设备，Ceveron(R)MFU-500，Technoclone)。使用Zetasizer Nano S(马尔文仪器有限公司)来评估MP的尺寸和分散性，该仪器采用173度背向散射检测器和N5亚微米粒度分析仪(Beckman Coulter)，使用多角度测量(30.1度、62.6度和90度)。在所使用的仪器中，只有使用30.1度测量角度的N5才能检测到预期粒度分布的MP，并演示通过过滤去除它们。MPS(平均粒径范围：Pre，101-464 nm；POST，21-182nmFilter)在过滤后显著减小(P<；0.0001)，但多分散性指数(中位数：Pre，0.746，Post，0.769)无明显变化。O型血样(Pre，247 nm)和A血型(Pre，289 nm)的MPS大小无显著差异(P=0.44)。DLS为评估Mp的大小和分布提供了一种新的方法，该技术可以很容易地被用作评估FFP或其他血液制品中MPS的手段。
We have previously shown that fresh-frozen plasma (FFP) contains red blood cell-derived procoagulant microparticles (MPs) that are removable by 0.2 mu m filtration. Given the limitations of current methods for accurately sizing MPs, we have applied the novel approach of dynamic light scattering (DLS) to characterize the size distributions of these MPs within FFP.Fresh-frozen plasma was prepared from blood Group A and O donations (n = 10 of each) after an overnight hold of whole blood at 4 degrees C. On the day of analysis, plasma was thawed to 37 degrees C and daughter aliquots were studied pre- and post-filtration (0.2 mu m filtration device, Ceveron (R) MFU-500, Technoclone). MP size and dispersity was assessed using a Zetasizer Nano S (Malvern Instruments Ltd), which employs a 173 degrees backscatter detector and an N5 Submicron Particle Size Analyser (Beckman Coulter) using multi-angle measurements (30.1 degrees, 62.6 degrees and 90 degrees). The analysers presented MP size distribution graphically as intensity plots, mean size, standard deviation and polydispersity index.Of the instruments used, only the N5 utilizing a 30.1 degrees angle of measurement could detect MPs of the expected size distribution and demonstrate their removal by filtration. MPs (range of mean particle diameters: pre, 101-464 nm; post, 21-182 nm filtration) were significantly smaller post-filtration (P < 0.0001), but polydispersity index (median: pre, 0.746, post, 0.769) exhibited no significant change. There was no significant difference between the size of MPs from blood Group O (pre, 247 nm) and Group A (pre, 289 nm) samples (P = 0.44).Our data demonstrates that DLS offers a novel approach to assessing MP size and distribution, a technique that could be easily adopted as a means of assessing MPs within either FFP or other blood products.