Impact of Channel Geometry on the Discrimination of Mechanically Impaired Red Blood Cells in Passive Microfluidics

Amirouche, Amin; Ferrigno, Rosaria; Faivre, Magalie

doi:10.3390/proceedings1040512

Cited by 4 publications

(4 citation statements)

References 6 publications

(5 reference statements)

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“…Pipetting results in shear of RBCs, which leads to the formation and fixation of flow shapes like knizocytes (also referred to as trilobes) (Lanotte et al, 2016), known to form at high shear rates (Figure 5). This highlights that the speed of fixation is extremely fast, knowing that RBCs in flow relax to the static shape in between 100 ms (Amirouche et al, 2017) and 1 s (Braunmüller et al, 2011). We observed knizocytes when fixing with 1% glutaraldehyde, but not when fixing with lower concentrations, hinting to a slower fixation for lower concentrations.…”

Section: Discussionmentioning

confidence: 97%

Glutaraldehyde – A Subtle Tool in the Investigation of Healthy and Pathologic Red Blood Cells

et al. 2019

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Glutaraldehyde is a well-known substance used in biomedical research to fix cells. Since hemolytic anemias are often associated with red blood cell shape changes deviating from the biconcave disk shape, conservation of these shapes for imaging in general and 3D-imaging in particular, like confocal microscopy, scanning electron microscopy or scanning probe microscopy is a common desire. Along with the fixation comes an increase in the stiffness of the cells. In the context of red blood cells this increased rigidity is often used to mimic malaria infected red blood cells because they are also stiffer than healthy red blood cells. However, the use of glutaraldehyde is associated with numerous pitfalls: (i) while the increase in rigidity by an application of increasing concentrations of glutaraldehyde is an analog process, the fixation is a rather digital event (all or none); (ii) addition of glutaraldehyde massively changes osmolality in a concentration dependent manner and hence cell shapes can be distorted; (iii) glutaraldehyde batches differ in their properties especially in the ratio of monomers and polymers; (iv) handling pitfalls, like inducing shear artifacts of red blood cell shapes or cell density changes that needs to be considered, e.g., when working with cells in flow; (v) staining glutaraldehyde treated red blood cells need different approaches compared to living cells, for instance, because glutaraldehyde itself induces a strong fluorescence. Within this paper we provide documentation about the subtle use of glutaraldehyde on healthy and pathologic red blood cells and how to deal with or circumvent pitfalls.

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Section: Discussionmentioning

confidence: 97%

Glutaraldehyde – A Subtle Tool in the Investigation of Healthy and Pathologic Red Blood Cells

et al. 2019

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“…This assumption would explain why at low hydrodynamic stress -where the cell properties dominate the recovery process and where the hydrodynamics can be neglected -we can assume being in an almost static configuration, the recovery time tend toward relaxation time values. In a previous study, we reported that at fixed hydrodynamic stress, shape recovery times of RBCs can be used to discriminate between healthy and mechanically impaired RBCs (Amirouche et al, 2017). In the present paper, we aim at demonstrating that our approach is sensitive enough to discriminate between different membrane modifications.…”

Section: Typical Behavior Of Healthy Rbcsmentioning

confidence: 80%

Mechanical Signature of Red Blood Cells Flowing Out of a Microfluidic Constriction Is Impacted by Membrane Elasticity, Cell Surface-to-Volume Ratio and Diseases

et al. 2020

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Despite the fact that Red Blood Cells (RBCs) have been intensively studied in the past 50 years to characterize mechanical phenotypes associated with both healthy and pathological states, only ektacytometry (i.e., laser diffractometry) is currently used by hematologists to screen for RBC membrane disorders. Therefore, the development of new diagnostic tools able to perform analysis at the scale of a single cell, over a statistically relevant population, would provide important complementary information. But these new diagnostic tools would have to be able to discriminate between different disorders causing a change in RBCs mechanical properties. We evaluated the mechanical response of artificially rigidified RBCs flowing through a microfluidic constriction. The geometry consists in a 50 µm wide channel with a succession of 14 tooth-like patterns, each composed of a 5 µm wide and 10 µm long constriction, associated with a 25 µm wide and 10 µm long enlargement. RBCs deformability was altered using two chemical treatments, known to affect RBCs membrane surface area and membrane deformability, lysolecithine (LPC) and diamide, respectively. Differences between samples were highlighted by the representation of the inverse of the shape recovery time (1/τ r ), versus the extension at the exit of the constriction, D out . The results demonstrate that our approach is able to provide a direct signature of RBCs membrane composition and architecture, as it allows discriminating the effect of changes in RBCs membrane surface area from changes in RBCs membrane deformability. Finally, in order to evaluate the potential of our microsystem to detect pathological cells, we have performed preliminary experiments on patients with Hereditary Spherocytosis (HS) or Sickle Cell Anemia (SCA).

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“…Mathematical models of deformable RBC with Discrete Element Methods have also been taken into account, along with the lattice Boltzmann models of immersed boundaries [172,173]. Other relevant studies include the assessment of shear dependencies in RBC adhesion [174], analysis of the changes in RBC stiffness [175], mechanical characterization of stored RBCs via mathematical models [176], donor-dependent aging curves based on microfluidic RBC models [177], effects of channel geometry in RBC sorting [178], and the assessment of RBC deformity using iron-dextran tests [179].…”

Section: Deformation Of Single Erythrocytes In Microchannelsmentioning

confidence: 99%

Advances in Microfluidics for Single Red Blood Cell Analysis

et al. 2023

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The utilizations of microfluidic chips for single RBC (red blood cell) studies have attracted great interests in recent years to filter, trap, analyze, and release single erythrocytes for various applications. Researchers in this field have highlighted the vast potential in developing micro devices for industrial and academia usages, including lab-on-a-chip and organ-on-a-chip systems. This article critically reviews the current state-of-the-art and recent advances of microfluidics for single RBC analyses, including integrated sensors and microfluidic platforms for microscopic/tomographic/spectroscopic single RBC analyses, trapping arrays (including bifurcating channels), dielectrophoretic and agglutination/aggregation studies, as well as clinical implications covering cancer, sepsis, prenatal, and Sickle Cell diseases. Microfluidics based RBC microarrays, sorting/counting and trapping techniques (including acoustic, dielectrophoretic, hydrodynamic, magnetic, and optical techniques) are also reviewed. Lastly, organs on chips, multi-organ chips, and drug discovery involving single RBC are described. The limitations and drawbacks of each technology are addressed and future prospects are discussed.

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Impact of Channel Geometry on the Discrimination of Mechanically Impaired Red Blood Cells in Passive Microfluidics

Cited by 4 publications

References 6 publications

Glutaraldehyde – A Subtle Tool in the Investigation of Healthy and Pathologic Red Blood Cells

Glutaraldehyde – A Subtle Tool in the Investigation of Healthy and Pathologic Red Blood Cells

Mechanical Signature of Red Blood Cells Flowing Out of a Microfluidic Constriction Is Impacted by Membrane Elasticity, Cell Surface-to-Volume Ratio and Diseases

Advances in Microfluidics for Single Red Blood Cell Analysis

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