Lavi Edwin D, Crivellari Francesca, Gagnon Zachary
Department of Chemical Engineering, Texas A&M University, College Station, Texas, USA.
Electrophoresis. 2022 Jun;43(12):1297-1308. doi: 10.1002/elps.202100241. Epub 2022 May 1.
The ability to transport and store a large human blood inventory for transfusions is an essential requirement for medical institutions. Thus, there is an important need for rapid and low-cost characterization tools for analyzing the properties of human red blood cells (RBCs) while in storage. In this study, we investigate the ability to use dielectrophoresis (DEP) for measuring the storage-induced changes in RBC electrical properties. Fresh human blood was collected, suspended in K2-EDTA anticoagulant, and stored in a blood bank refrigerator for a period of 20 days. Cells were removed from storage at 5-day intervals and subjected to a glutaraldehyde crosslinking reaction to "freeze" cells at their ionic equilibrium at that point in time and prevent ion leakage during DEP analysis. The DEP behavior of RBCs was analyzed in a high permittivity DEP buffer using a three-dimensional DEP chip (3DEP) and also compared to measurements taken with a 2D quadrupole electrode array. The DEP analysis confirms that RBC electrical property changes occur during storage and are only discernable with the use of the cell crosslinking reaction above a glutaraldehyde fixation concentration of 1.0 w/v%. In particular, cytoplasm conductivity was observed to decrease by more than 75% while the RBC membrane conductance was observed to increase by more than 1000% over a period of 20 days. These results show that the presented combination of chemical crosslinking and DEP can be used as rapid characterization tool for monitoring electrical properties changes of human RBCs while subjected to refrigeration in blood bank storage.
对于医疗机构而言,具备运输和储存大量用于输血的人类血液库存的能力是一项基本要求。因此,迫切需要快速且低成本的表征工具,用于分析储存过程中人类红细胞(RBC)的特性。在本研究中,我们探究了利用介电电泳(DEP)测量储存诱导的RBC电学特性变化的能力。采集新鲜人类血液,悬浮于K2 - EDTA抗凝剂中,并在血库冰箱中储存20天。每隔5天从储存样本中取出细胞,使其进行戊二醛交联反应,以便在该时间点将细胞“固定”在其离子平衡状态,并防止在DEP分析过程中离子泄漏。使用三维DEP芯片(3DEP)在高介电常数DEP缓冲液中分析RBC的DEP行为,并与二维四极电极阵列的测量结果进行比较。DEP分析证实,RBC电学特性在储存过程中会发生变化,并且只有在戊二醛固定浓度高于1.0 w/v%时使用细胞交联反应才能辨别这些变化。特别是,在20天的时间里,观察到细胞质电导率下降超过75%,而RBC膜电导率增加超过1000%。这些结果表明,所提出的化学交联和DEP的组合可作为一种快速表征工具,用于监测血库储存中冷藏条件下人类RBC电学特性的变化。