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直接从全血中并行免疫磁珠分离嗜碱性粒细胞

Parallelized Immunomagnetic Isolation of Basophils Directly from Whole Blood.

作者信息

Myles Justin, Castaño Nicolas, Kim Sungu, Zhu Zhenyun, Tang Sindy K Y

机构信息

Department of Mechanical Engineering, Stanford University, Stanford, CA 94305, USA.

出版信息

Adv Nanobiomed Res. 2024 Feb;4(2). doi: 10.1002/anbr.202300122. Epub 2023 Dec 3.

Abstract

Basophils are the rarest circulating white blood cells (WBCs), but they play important roles in allergic disorders and other diseases. To enhance diagnostic capabilities, it would be desirable to isolate and analyze basophils efficiently from small blood samples. In 100 μL of whole blood, there are typically 10 basophils, outnumbered by ~10 WBCs and ~10 red blood cells (RBCs). Basophils' low abundance has therefore presented a significant challenge in their isolation from whole blood. Conventional in-bulk basophil isolation methods require lengthy processing steps and cannot work with small volumes of blood. Here we report a parallelized integrated basophil isolation device (pi-BID) for the negative immunomagnetic selection of basophils directly from 4 samples of 100 μL of whole blood, in parallel, within 14 minutes including sample preparation time. The pi-BID interfaces directly with standard sample tubes, and uses a single pressure source to drive the flow in parallel microfluidic channels. Compared with conventional in-bulk basophil isolation, the pi-BID is >3× faster, and has higher purity (93%) and similar recovery (~67%). Compared with other microfluidic devices for the immunomagnetic isolation of WBC sub-types, our pi-BID achieves 10× higher enrichment of target cells from whole blood, with no prior removal of RBCs necessary.

摘要

嗜碱性粒细胞是循环系统中最为罕见的白细胞,但它们在过敏性疾病及其他病症中发挥着重要作用。为提高诊断能力,期望能从小血样中高效分离并分析嗜碱性粒细胞。在100微升全血中,通常约有10个嗜碱性粒细胞,其数量比约10个白细胞和10个红细胞少。因此,嗜碱性粒细胞的低丰度给从全血中分离它们带来了重大挑战。传统的批量嗜碱性粒细胞分离方法需要冗长的处理步骤,且无法处理少量血液。在此,我们报告了一种并行集成嗜碱性粒细胞分离装置(pi - BID),用于直接从4份100微升全血样本中并行进行嗜碱性粒细胞的阴性免疫磁选,包括样本制备时间在内,整个过程在14分钟内完成。pi - BID可直接与标准样本管连接,并使用单一压力源驱动并行微流控通道中的流体。与传统的批量嗜碱性粒细胞分离方法相比,pi - BID速度快3倍以上,纯度更高(约93%),回收率相似(约67%)。与其他用于白细胞亚型免疫磁分离的微流控装置相比,我们的pi - BID从全血中富集靶细胞的效率提高了10倍,且无需事先去除红细胞。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b818/11244651/990ff1c1b1b5/nihms-1952772-f0001.jpg

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