Size and density measurements of single sickle red blood cells using microfluidic magnetic levitation.
Size and density measurements of single sickle red blood cells using microfluidic magnetic levitation.
复制标题
DOI：
10.1039/d1lc00686j
复制
发表时间：
2022-02-15
期刊：
影响因子：
6.1
通讯作者：
中科院分区：
文献类型：
--
作者：
单细胞具有独特的生物物理特征，在各种健康和病理状态下可以迅速改变。例如，细胞密度是细胞类型之间不同的固有属性。表征下至单细胞水平的基本密度特性的戏剧性变化可以揭示处于病理状态的亚群。在这里，我们开发了一种基于微流控、磁悬浮的分析(MagDense)，它可以检测单个红细胞(RBC)的微小密度差异，分辨率高达0.0001克/毫升。这种分析方法根据红细胞在非离子顺磁介质中的密度分布对红细胞进行分级，该介质流动在放置在相同磁极相互面对的磁铁之间的毛细管微通道中。基于单个红细胞在特定平衡状态下的悬浮高度和密度的精确测量，我们证明了MagDense可以准确地分析含HBS和含HBA的红细胞的密度。此外，单细胞水平的精确密度和细胞大小测量显示，在接受输血的纯合子镰状细胞病患者的血液样本中，有三种不同的红细胞亚群；密度较低的含HBA的红细胞悬浮较高，而密度较高的含HBS的红细胞悬浮较低。我们比较了镰刀状细胞病受试者和非镰状细胞病对照组的平均红细胞密度，发现每组患者的红细胞密度密度带明显分开，表示血液样本中细胞密度的可能范围。我们方法的高分辨率使测量与平均红细胞密度的偏差成为可能。此外，我们还引入了一个新的术语作为密度分散的度量，即RBC漂浮密度宽度。镰状细胞病的平均红细胞密度与全血血红蛋白相关(p=0.032，直线回归)，红细胞平均密度与网织红细胞绝对数和全血红细胞分布宽度相关(p=0.002和p=0.002，直线回归)。我们基于磁悬浮的检测方法能够快速、准确、基于密度地对单个红细胞进行成像、分析和无标记监测。我们的方法可以广泛应用于研究血细胞疾病和新出现的治疗方法对患者预后的影响。
Single cells have unique biophysical signatures that can rapidly change during various healthy and pathological states. For instance, cellular density is an inherent property differing between cell types. Characterizing dramatic changes in fundamental density properties down to the single-cell level can reveal sub-populations in pathological states. Here, we have developed a microfluidic, magnetic levitation-based assay (MagDense) that detects minute density differences of individual red blood cells (RBCs) down to 0.0001 g/mL resolution. This assay fractionates RBCs based on their density profiles in a non-ionic paramagnetic medium flowing in a capillary microchannel placed between magnets with same poles facing each other. Based on precisely measured levitation height and density of individual RBCs at their specific equilibrium state, we demonstrated that MagDense can accurately analyze the density of HbS-containing RBCs and HbA-containing RBCs. In addition, the precise density and cell size measurements at the single cell level showed three different sub-populations of RBCs in blood samples from individuals with homozygous sickle cell disease receiving blood transfusions; where less dense, HbA-containing RBCs levitated higher, while the denser, HbS-containing RBCs levitated lower. We compared the mean RBC densities of sickle cell disease subjects with non-sickle controls and found distinctly separated density bands of RBC density for each group denoting the likely range of cell densities seen in the blood samples. The high resolution of our method enabled measurement of deviation from the mean RBC density. Moreover, we introduced a new term as a measure of density dispersion, “RBC Levitational Density Width, RLDW”. Mean RBC density in sickle cell disease associated with hemoglobin from complete blood count (p=0.032, Linear regression) and RLDW associated with absolute reticulocyte count (ARC) and RBC distribution width (RDW) from complete blood count (p=0.002 for ARC and p=003 for RDW, linear regression). Our magnetic levitation-based assay enables rapid, accurate, density-based imaging, profiling and label-free monitoring of single RBCs. Our approach can be broadly applicable to investigate blood cell disorders and the effects of emerging curative therapies in patient outcomes.
登录
查看更多内容
影响因子：
6.1
作者：
Bryan AK;Hecht VC;Shen W;Payer K;Grover WH;Manalis SR
通讯作者：
Manalis SR
影响因子：
6.5
作者：
HORIUCHI, K;STEPHENS, MJ;OHENEFREMPONG, K
通讯作者：
OHENEFREMPONG, K
影响因子：
20.3
作者：
Bartolucci, Pablo;Brugnara, Carlo;Galacteros, Frederic
通讯作者：
Galacteros, Frederic
影响因子：
2.4
作者：
Kaul, DK;Fabry, ME
通讯作者：
Fabry, ME
影响因子：
4
作者：
Lew VL;Tiffert T
通讯作者：
Tiffert T