Label-Free and Noninvasive Single-Cell Characterization for the Viscoelastic Properties of Cryopreserved Human Red Blood Cells Using a Dielectrophoresis-On-a-Chip Approach

Abstract

Successful human red blood cell cryopreservation techniques have been gradually developed in recent decades, with great potential for use in clinical medicine and basic research. The mechanical properties of a single cell are important clues to reveal the physiological and pathological state of the red blood cell, but they have not been used to assess the physiological state of the cell after cryopreservation. Herein, we investigated the effects of cryopreservation processes on human red blood cell biomechanics by means of a microfluidic, label-free, synchronous, and nondestructive electrodeformation assay. We found that the effects of viscoelasticity of the red blood cell membrane between permeable and impermeable cryoprotectants were different. Our findings showed that high freeze−thaw recovery did not mean that the recovered cells had excellent viscoelasticity. The results demonstrate that single-cell viscoelasticity is an irreplaceable indicator for assessing the quality of the recovered cells and help us to deepen understanding of the cryoinjury mechanism. Assessment of the single cell viscoelasticity offers significant potential for application in optimizing the cryopreservation process and screening optimal cryoprotectants.

Advances in Microfluidics for Single Red Blood Cell Analysis

“…Technologies based on the dielectrophoresis effects are effective for the analysis of fatigue, force, and stress at the cellular level. For example, cell manipulation systems have been widely employed to measure cellular biomechanics using microfluidic platforms, including studies on cell stretching and manipulation [135,136], electrical property changes of stored RBC [137], label-free and noninvasive characterization for the viscoelastic properties of RBC [138],benchmarking dielectrophoretic separation metrics of unknown types of RBC (healthy, modified, …) [139], the oxidative stress analysis for RBCs (Figure 9A) [140], dynamic fatigue measurements [141], detecting circadian rhythms in RBCs [142], nonlinear viscoelastic analyses (Figure 9B) [143], biomechanics of erythrocyte membrane failures [144], liquid metal electrode-based dielectrophoretic schemes [145], and a portable system with multiple dielectrophoretic applications for RBC analyses [146].…”

“…Technologies based on the dielectrophoresis effects are effective for the analysis of fatigue, force, and stress at the cellular level. For example, cell manipulation systems have been widely employed to measure cellular biomechanics using microfluidic platforms, including studies on cell stretching and manipulation [135,136], electrical property changes of stored RBC [137], label-free and noninvasive characterization for the viscoelastic properties of RBC [138], benchmarking dielectrophoretic separation metrics of unknown types of RBC (healthy, modified, . .…”

A Dynamic Membrane‐Active Glycopeptide for Enhanced Protection of Human Red Blood Cells against Freeze‐Stress

“…[7][8][9][10][11] It is clear, hence, that there exist two different methods for measuring the cryoprotective outcome, the cryosurvival (the cell count of RBCs postthaw to total cells initially frozen) [6,7,9] and the cell recovery post-thaw (the cell count of RBCs post-thaw to total fresh cells initially incubated). [12][13][14] Currently, it is favorable to deliver appropriate trehalose into RBCs by membrane-perturbative compounds and subsequently perform direct cryostorage to achieve both the excellent cryoprotective outcomes, which is still challenging.…”

High-throughput and directed microparticle manipulation in complex-shaped maze chambers based on travelling surface acoustic waves

“…6,7 The simultaneous manipulation of high-throughput microparticles along complex trajectories has become an urgent interdisciplinary demand in numerous applications. 8–10 Several techniques have been proposed and employed to manipulate multiple microparticles, including optical tweezers, 11–13 electrical tweezers, 14–16 magnetic tweezers, 17,18 and acoustic tweezers. 19,20 Optical and electrical tweezers can manipulate multiple microparticles at the same time.…”