Abstract:Changes in red blood cell (RBC) deformability are associated with the pathology of many diseases and could potentially be used to evaluate disease status and treatment efficacy. We developed a simple, sensitive, and multiplexed RBC deformability assay based on the spatial dispersion of single cells in structured microchannels. This mechanism is analogous to gel electrophoresis, but instead of transporting molecules through nano-structured material to measure their length, RBCs are transported through micro-str… Show more
“…Traditional single cell manipulation, including micropipette aspiration 33,34 , optical tweezers 35 , and atomic force microscopy 36 , involve technically challenging experiments that require skilled personnel and highly specialized equipment. Microfluidic approaches for deformation based RBC deformability analysis include the measurement of capillary obstruction 37 , wedging in tapered constrictions 38,39 , transiting time through constrictions [40][41][42][43] , and transiting pressure through constrictions 17,44,45 . While these approaches greatly simplified RBC deformability analysis, the throughput of these processes are still relatively limited.…”
A fundamental challenge in the transfusion of red blood cells (RBCs) is that a subset of donated RBC units may not provide optimal benefit to transfusion recipients. This variability stems from the inherent ability of donor RBCs to withstand the physical and chemical insults of cold storage, which ultimately dictate their survival in circulation. The loss of RBC deformability during cold storage is well-established and has been identified as a potential biomarker for the quality of donated RBCs. While RBC deformability has traditionally been indirectly inferred from rheological characteristics of the bulk suspension, there has been considerable interest in directly measuring the deformation of RBCs. Microfluidic technologies have enabled single cell measurement of RBC deformation but have not been able to consistently distinguish differences between RBCs between healthy donors. Using the microfluidic ratchet mechanism, we developed a method to sensitively and consistently analyze RBC deformability. We found that the aging curve of RBC deformability varies significantly across donors, but is consistent for each donor over multiple donations. Specifically, certain donors seem capable of providing RBCs that maintain their deformability during two weeks of cold storage in standard test tubes. The ability to distinguish between RBC units with different storage potential could provide a valuable opportunity to identify donors capable of providing RBCs that maintain their integrity, in order to reserve these units for sensitive transfusion recipients.Donor-dependent Red Cell Aging
“…Traditional single cell manipulation, including micropipette aspiration 33,34 , optical tweezers 35 , and atomic force microscopy 36 , involve technically challenging experiments that require skilled personnel and highly specialized equipment. Microfluidic approaches for deformation based RBC deformability analysis include the measurement of capillary obstruction 37 , wedging in tapered constrictions 38,39 , transiting time through constrictions [40][41][42][43] , and transiting pressure through constrictions 17,44,45 . While these approaches greatly simplified RBC deformability analysis, the throughput of these processes are still relatively limited.…”
A fundamental challenge in the transfusion of red blood cells (RBCs) is that a subset of donated RBC units may not provide optimal benefit to transfusion recipients. This variability stems from the inherent ability of donor RBCs to withstand the physical and chemical insults of cold storage, which ultimately dictate their survival in circulation. The loss of RBC deformability during cold storage is well-established and has been identified as a potential biomarker for the quality of donated RBCs. While RBC deformability has traditionally been indirectly inferred from rheological characteristics of the bulk suspension, there has been considerable interest in directly measuring the deformation of RBCs. Microfluidic technologies have enabled single cell measurement of RBC deformation but have not been able to consistently distinguish differences between RBCs between healthy donors. Using the microfluidic ratchet mechanism, we developed a method to sensitively and consistently analyze RBC deformability. We found that the aging curve of RBC deformability varies significantly across donors, but is consistent for each donor over multiple donations. Specifically, certain donors seem capable of providing RBCs that maintain their deformability during two weeks of cold storage in standard test tubes. The ability to distinguish between RBC units with different storage potential could provide a valuable opportunity to identify donors capable of providing RBCs that maintain their integrity, in order to reserve these units for sensitive transfusion recipients.Donor-dependent Red Cell Aging
“…However, both methods are time and labor intensive (tens of cells per hour), posing challenges for examining large populations of cells to either obtain statistically valid conclusions or identify rare sub-populations. Recent advances in micro-/nano-fabrication technologies have opened up a range of new mechanophenotyping technologies that can measure deformations of tens to hundreds of cells per second [11][12][13][14][15][16][17][18][19] . We recently reported a technology, called deformability cytometry, in which a cross-slot microfluidic channel is employed to generate a hydrodynamic extension zone where individual cells are exposed to uniform hydrodynamic stress and deformed 20 .…”
In this report, we present multiparameter deformability cytometry (m-DC), in which we explore a large set of parameters describing the physical phenotypes of pluripotent cells and their derivatives. m-DC utilizes microfluidic inertial focusing and hydrodynamic stretching of single cells in conjunction with high-speed video recording to realize high-throughput characterization of over 20 different cell motion and morphology-derived parameters. Parameters extracted from videos include size, deformability, deformation kinetics, and morphology. We train support vector machines that provide evidence that these additional physical measurements improve classification of induced pluripotent stem cells, mesenchymal stem cells, neural stem cells, and their derivatives compared to size and deformability alone. In addition, we utilize visual interactive stochastic neighbor embedding to visually map the high-dimensional physical phenotypic spaces occupied by these stem cells and their progeny and the pathways traversed during differentiation. This report demonstrates the potential of m-DC for improving understanding of physical differences that arise as cells differentiate and identifying cell subpopulations in a label-free manner. Ultimately, such approaches could broaden our understanding of subtle changes in cell phenotypes and their roles in human biology.
“…4(d)). 61 Based on application of di®erent micro°uidic channels, they all suggested that erythrocyte deformability decreased signi¯cantly under various abnormal conditions, such as malaria-infection, 62,63 blood storage lesion, 64,65 diabetes 66,67 and sickle cell disease. 68,69 Compared with the optical tweezers and the micropipette aspiration, the micro°uidic channels provided a unique opportunity to investigate erythrocyte deformability in a rapid, highthroughput and relatively easy-to-use method at single-cell level.…”
As the indispensable oxygen-transporting cells, erythrocytes exhibit extreme deformability and amazing stability as they are subject to huge reversible shear stress and extrusion force during massive circulation in the body. The unique architecture of spectrin-actin-based membrane-skeleton is considered to be responsible for such excellent mechanical properties of erythrocytes. Although erythrocytes have been recognized for more than 300 years, myriad questions about membrane-skeleton constantly attract people’s attention. Here, we summarize the kinds of distinctive single-cell and single-molecule techniques that were used to investigate the structure and function of erythrocyte membrane-skeleton at macro and micro levels.
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