Research progress on mechanical and flow properties of blood cells in microcirculation using microfluidic devices
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摘要: 近年来，随着微流控芯片技术的快速发展，微流控芯片在生物医学研究领域得到了广泛关注。由于其具有高通量、高灵敏度、集成化、低消耗及可控化等诸多特点，为在多细胞水平研究细胞迁移和分选动力学提供了新的技术平台。利用微流控芯片微通道结构设计灵活的特点，可在实验条件下模拟正常的生理和病理条件下的复杂血管；其微米尺寸的微通道也适于单细胞引入、操纵及检测。因此，用微流控芯片技术在单细胞层面对细胞生物力学性能表征也引起了广泛关注。以健康和疾病中的血细胞为例，从单细胞变形、流动、黏附、机械疲劳等力学性能表征到多细胞迁移及分离动力学等方面归纳目前微流控芯片技术在细胞力学分析和表征方面的研究进展。Abstract: Microfluidics (or lab-on-a-chip) is an important technology suitable for a wide range of biomedical applications from single-cell analysis to point-of-care diagnosis. In this paper, we review recent advances in the applications of the microfluidic technology in the field of cell biology and biomechanics. We highlight examples of some successful applications of microfluidic devices in probing the mechanical and rheological characteristics of blood cells in healthy and diseased states at single-cell and multi-cell levels, and in investigating the cell migration and separation at the whole-cell population.
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Key words:
- micro/nano flow /
- blood cell /
- cell mechanics /
- cell dynamics /
- cell separation
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图 1 血细胞在血管内流动的动力学特征及微流控仿生模型设计
Figure 1. The hydrodynamics of blood flow and its microfluidic culture models
图 2 基于单通道微流控装置研究健康红细胞在毛细管内的流动和变形[10]。(a)~(d)显示红细胞在流过毛细管不同区域时的变形
Figure 2. In vitro microfluidic model for investigating the translocation process of a healthy human red blood cell (RBC) through a microcapillary[10]. The series of images (a)~(d) show the shape of the RBC as it is squeezed through the narrow microchannel
图 3 基于单通道微流控装置研究疟疾感染红细胞在不同尺寸的毛细管内的流动及堵塞行为[14]。自左至右毛细管的宽度分别为8、6、4及2 μm。感染初期的环状体红细胞可顺利通过毛细管道((a1)~(a4)); 感染末期处于裂殖体阶段的红细胞仅能通过8 μm的毛细管道(b1), 但无法通过6 μm及以下的毛细管并导致微通道的堵塞((b2)~(b4))
Figure 3. Micrographs of malaria-infected RBCs traversal across microfluidic channels. Ring-stage infected RBCs can pass through all constricted microchannels ((a1)~(a4)); schizont-stage infected RBCs block all but the 8 μm microchannel ((b1)~(b4))
图 4 基于单通道"漏斗形收缩"微流控装置研究疟疾感染红细胞的流动及变形行为[15]。(a)感染初期的环状体红细胞在较小的压力梯度下可通过漏斗形收缩微管道; (b)感染末期处于裂殖体阶段的红细胞则需要较大的压力梯度才能通过微细管; (c)基于实验结果统计得到的不同感染阶段的红细胞膜皮质张力
Figure 4. Analysis of the flow behavior and deformability of malaria-infected RBCs in microfluidic channels with multiple funnel-shaped constrictions. (a) A ring-stage infected RBC could transit rapidly through the funnel constriction at a low pressure; (b) A schizont-stage infected RBC, which is hardly deformed, requires a high pressure to drive through the constriction; (c) Histogram of the measured cortical tension of uninfected and malaria-infected RBCs
图 5 基于多通道微流控装置研究受疟疾感染红细胞在毛细管内的流动行为[17]。(a)微流道装置示意图。微流道内有多组并排的宽度仅为3 μm的微管道阵列, 红细胞流过微管道阵列时需变形才能通过; (b)通过实验获得的健康红细胞(蓝色箭头所示)和疟疾感染红细胞(红色箭头所示)在多通道毛细管内的流动现象
Figure 5. Microfluidic platform for studying the flow behavior of malaria-infected RBCs in the microchannel[17]. (a) Schematic diagram of the device design; the spacing between pillars is 3 μm and the depth of the device is 4.2 μm; (b) Micrographs of uninfected (blue arrows) and malaria-infected (red arrows) RBCs in the microchannel
图 6 基于多通道微流控装置研究镰状细胞贫血症下红细胞的流动和变形性能[18]。(a)通过调控氧气水平和脱氧程度控制红细胞的镰变和可逆转镰变过程; (b)当处于缺氧状态时, 红细胞发生镰变后导致其形状发生变化(黄色箭头所示); (c)微流道装置示意图。微流道内有多组并排的宽度仅为4 μm的微通道阵列, 红细胞需变形才能通过; (d)在有氧(上图)和脱氧条件(下图)下, 观察红细胞在毛细管道内的流动及堵塞现象
Figure 6. Microfluidic platform for studying sickle cell behavior under transient hypoxic conditions[18]. (a) Schematic diagram of microfluidic device; (b) Visual determination of cell sickling events from morphological changes in RBC membrane; (c) Schematic diagram of microfluidic device; the microfluidic channel contains periodic obstacles forming 15-μm-long, 4-μm-wide and 5-μm-high microgates; (d) Individual RBC behavior under oxygenated (Oxy, upper) and deoxygenated (DeOxy, lower) states[18]
图 7 基于多通道微流控装置模拟微血管堵塞及血栓形成过程[25]。(a)微流控芯片装置图; (b)微流控芯片中多通路微管道网络示意图, 其中最中间一排的微管道宽度只有30 μm; (c)微流控芯片上"内皮细胞化"微血管分叉结构; (d)血细胞-内皮细胞黏附及多细胞聚集导致的微血管堵塞现象
Figure 7. In vitro microfluidic microvasculature model for modeling of the microvascular occlusion and thrombosis that occur in hematologic disease[25]. (a)~(b) Illustration of the device design; (c) Characterization of the "endothelialized" microvasculature on the chip; (d) The observation of microchannel occlusion due to a combination of increased adhesion and cell stiffness
图 8 基于惯性效应和黏弹性效应设计的微流控装置用于特定类型血细胞的分离采集[34-35]。(a)微流道内疟疾感染红细胞和健康红细胞的分离[34]; (b)微流道内循环肿瘤细胞、白细胞和红细胞的分离[35]
Figure 8. Schematic illustration of cell separation in microfluidic channels based on cell size and deformability [34-35]. (a) Separation of malaria-infected RBCs and uninfected RBCs in microfluidic channel [34]; (b) Separation of RBCs, white blood cells (WBCs) and tumor cells in microfluidic channel[35]
图 9 微流控通道中细胞分离示意图。(a)~(b)基于惯性升力和磁场效应从微量血液样品中实现肿瘤细胞和血细胞的分离采集[30, 41]; (c)~(d)通过超声波微流控装置从血细胞混合液中分离肿瘤细胞[43-45]
Figure 9. Schematic illustration of cell separation in microfluidic channel. (a)~(b) Separation of blood cells and tumor cells in ferrofluids[30, 41]; (c)~(d) acoustic separation of tumor cells from blood samples[43-45]
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