Mechanical Signature of Red Blood Cells Flowing Out of a Microfluidic Constriction Is Impacted by Membrane Elasticity, Cell Surface-to-Volume Ratio and Diseases
Mechanical Signature of Red Blood Cells Flowing Out of a Microfluidic Constriction Is Impacted by Membrane Elasticity, Cell Surface-to-Volume Ratio and Diseases
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DOI：
10.3389/fphys.2020.00576
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发表时间：
2020-06-12
影响因子：
4
通讯作者：
Connes, Philippe
中科院分区：
文献类型：
--
作者：
Faivre, Magalie;Renoux, Celine;Connes, Philippe
尽管在过去的50年里，人们已经对红细胞（RBC）进行了深入的研究，以表征与健康和病理状态相关的机械表型，但目前血液学家只使用细胞计数法（即激光衍射法）来筛查红细胞膜疾病。因此，新的诊断工具的开发能够在单个细胞的规模上进行分析，在统计相关的人群中，将提供重要的补充信息。但是这些新的诊断工具必须能够区分引起红细胞机械特性改变的不同疾病。我们评估了人工硬化红细胞流过微流体收缩的机械反应。几何形状由一个50 μ m宽的通道组成，其中有14个连续的齿状图案，每个图案由5 μ m宽和10 μ m长的收缩组成，并与25 μ m宽和10 μ m长的扩张相关联。红细胞的变形能力是通过两种化学处理来改变的，已知会影响红细胞的膜表面积和膜变形能力，分别是溶卵磷脂（LPC）和二胺。通过形状恢复时间的倒数（1/tau(r)）与收缩出口的延伸（out）的表示，突出了样本之间的差异。结果表明，我们的方法能够提供红细胞膜组成和结构的直接特征，因为它允许区分红细胞膜表面积变化的影响和红细胞膜可变形性变化的影响。最后，为了评估我们的微系统检测病理细胞的潜力，我们对遗传性球形细胞增生症（HS）或镰状细胞贫血症（SCA）患者进行了初步实验。
Despite the fact that Red Blood Cells (RBCs) have been intensively studied in the past 50 years to characterize mechanical phenotypes associated with both healthy and pathological states, only ektacytometry (i.e., laser diffractometry) is currently used by hematologists to screen for RBC membrane disorders. Therefore, the development of new diagnostic tools able to perform analysis at the scale of a single cell, over a statistically relevant population, would provide important complementary information. But these new diagnostic tools would have to be able to discriminate between different disorders causing a change in RBCs mechanical properties. We evaluated the mechanical response of artificially rigidified RBCs flowing through a microfluidic constriction. The geometry consists in a 50 mu m wide channel with a succession of 14 tooth-like patterns, each composed of a 5 mu m wide and 10 mu m long constriction, associated with a 25 mu m wide and 10 mu m long enlargement. RBCs deformability was altered using two chemical treatments, known to affect RBCs membrane surface area and membrane deformability, lysolecithine (LPC) and diamide, respectively. Differences between samples were highlighted by the representation of the inverse of the shape recovery time (1/tau(r)), versus the extension at the exit of the constriction,D-out. The results demonstrate that our approach is able to provide a direct signature of RBCs membrane composition and architecture, as it allows discriminating the effect of changes in RBCs membrane surface area from changes in RBCs membrane deformability. Finally, in order to evaluate the potential of our microsystem to detect pathological cells, we have performed preliminary experiments on patients with Hereditary Spherocytosis (HS) or Sickle Cell Anemia (SCA).