scholarly journals Linker histone H1 determines cell stiffness and differentiation

2020 ◽  
Author(s):  
Marie Kijima ◽  
Hiroyuki Yamagishi ◽  
Riho Kawano ◽  
Tomoki Konishi ◽  
Takuya Okumura ◽  
...  
Keyword(s):  
Lateral Displacement ◽  
Histone H1 ◽  
Cell Stiffness ◽  
Label Free ◽  
Size Fractionation ◽  
Mechanical Stiffness ◽  
Deterministic Lateral Displacement ◽  
Linear Relationships ◽  
Nuclear Changes ◽  
Positively Charged

ABSTRACTThe flexibility or stiffness, one mechanical property of cells, is a promising label-free biomarker for underlying cytoskeletal or nuclear changes associated with various disease processes and changes in cell state. However, the molecular changes responsible for the whole-cell mechanical stiffness remain to be clarified. Recently, it was shown that the deterministic lateral displacement (DLD) microfluidic device, originally developed for size fractionation of some particles, might be applied to distinguish cells according to their stiffness. In this experiment, using the DLD device and various cell lines differentially expressing histone H1, a positively-charged protein localized in the linker region of chromatin, we found linear relationships between histone H1 quantity and cell stiffness. We also found that the histone H1 quantity affected cell size and even cell differentiation.

scholarly journals Combining Electrostatic, Hindrance and Diffusive Effects for Predicting Particle Transport and Separation Efficiency in Deterministic Lateral Displacement Microfluidic Devices

Biosensors ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 126
Author(s):  
Valentina Biagioni ◽  
Giulia Balestrieri ◽  
Alessandra Adrover ◽  
Stefano Cerbelli
Keyword(s):  
Particle Transport ◽  
Separation Efficiency ◽  
Lateral Displacement ◽  
Operating Conditions ◽  
Separation Performance ◽  
Label Free ◽  
Deterministic Lateral Displacement ◽  
Promising Tool ◽  
Label Free Detection ◽  
Diffusive Effects

Microfluidic separators based on Deterministic Lateral Displacement (DLD) constitute a promising technique for the label-free detection and separation of mesoscopic objects of biological interest, ranging from cells to exosomes. Owing to the simultaneous presence of different forces contributing to particle motion, a feasible theoretical approach for interpreting and anticipating the performance of DLD devices is yet to be developed. By combining the results of a recent study on electrostatic effects in DLD devices with an advection–diffusion model previously developed by our group, we here propose a fully predictive approach (i.e., ideally devoid of adjustable parameters) that includes the main physically relevant effects governing particle transport on the one hand, and that is amenable to numerical treatment at affordable computational expenses on the other. The approach proposed, based on ensemble statistics of stochastic particle trajectories, is validated by comparing/contrasting model predictions to available experimental data encompassing different particle dimensions. The comparison suggests that at low/moderate values of the flowrate the approach can yield an accurate prediction of the separation performance, thus making it a promising tool for designing device geometries and operating conditions in nanoscale applications of the DLD technique.

scholarly journals Microfluidic label-free bioprocessing of human reticulocytes from erythroid culture

Lab on a Chip ◽  
2020 ◽  
Vol 20 (18) ◽  
pp. 3445-3460
Author(s):  
Kerwin Kwek Zeming ◽  
Yuko Sato ◽  
Lu Yin ◽  
Nai-Jia Huang ◽  
Lan Hiong Wong ◽  
...  
Keyword(s):  
Stem Cells ◽  
Hematopoietic Stem Cells ◽  
Closed Loop ◽  
Lateral Displacement ◽  
Label Free ◽  
Hematopoietic Stem ◽  
Deterministic Lateral Displacement ◽  
Dean Flow ◽  
Flow Fractionation ◽  
Human Rbcs

Developments in Dean flow fractionation (DFF) and deterministic lateral displacement (DLD) for label-free purification of cultured RBCs from human hematopoietic stem cells. An advancement in sorting and closed-loop manufacturing of viable human RBCs.

scholarly journals Separation of blood cells with differing deformability using deterministic lateral displacement

2014 ◽  
Vol 4 (6) ◽  
pp. 20140011 ◽  
Author(s):  
David Holmes ◽  
Graeme Whyte ◽  
Joe Bailey ◽  
Nuria Vergara-Irigaray ◽  
Andrew Ekpenyong ◽  
...  
Keyword(s):  
Whole Blood ◽  
Blood Cells ◽  
Lateral Displacement ◽  
Measurement Techniques ◽  
Cell Stiffness ◽  
Deterministic Lateral Displacement ◽  
Human Red Blood Cells ◽  
Hydrodynamic Behaviour ◽  
Novel Approach ◽  
Marker Free

Determining cell mechanical properties is increasingly recognized as a marker-free way to characterize and separate biological cells. This emerging realization has led to the development of a plethora of appropriate measurement techniques. Here, we use a fairly novel approach, deterministic lateral displacement (DLD), to separate blood cells based on their mechanical phenotype with high throughput. Human red blood cells were treated chemically to alter their membrane deformability and the effect of this alteration on the hydrodynamic behaviour of the cells in a DLD device was investigated. Cells of defined stiffness (glutaraldehyde cross-linked erythrocytes) were used to test the performance of the DLD device across a range of cell stiffness and applied shear rates. Optical stretching was used as an independent method for quantifying the variation in stiffness of the cells. Lateral displacement of cells flowing within the device, and their subsequent exit position from the device were shown to correlate with cell stiffness. Data showing how the isolation of leucocytes from whole blood varies with applied shear rate are also presented. The ability to sort leucocyte sub-populations (T-lymphocytes and neutrophils), based on a combination of cell size and deformability, demonstrates the potential for using DLD devices to perform continuous fractionation and/or enrichment of leucocyte sub-populations from whole blood.

scholarly journals On the Three-Dimensional Structure of the Flow through Deterministic Lateral Displacement Devices and Its Effects on Particle Separation

Processes ◽  
2019 ◽  
Vol 7 (8) ◽  
pp. 498 ◽  
Author(s):  
Valentina Biagioni ◽  
Alessandra Adrover ◽  
Stefano Cerbelli
Keyword(s):  
Lateral Displacement ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Periodic Lattice ◽  
Separation Mechanism ◽  
Label Free ◽  
Deterministic Lateral Displacement ◽  
Slip Conditions ◽  
Flow Through ◽  
The Impact

Experiments have shown that a suspension of particles of different dimensions pushed through a periodic lattice of micrometric obstacles can be sorted based on particle size. This label-free separation mechanism, referred to as Deterministic Lateral Displacement (DLD), has been explained hinging on the structure of the 2D solution of the Stokes flow through the patterned geometry, thus neglecting the influence of the no-slip conditions at the top and bottom walls of the channel hosting the obstacle lattice. We show that the no-slip conditions at these surfaces trigger the onset of off-plane velocity components, which impart full three-dimensional character to the flow. The impact of the 3D flow structure on particle transport is investigated by enforcing an excluded volume approach for modelling the interaction between the finite-sized particles and the solid surfaces. We find that the combined action of particle diffusion and of the off-plane velocity component causes the suspended particles to migrate towards the top and bottom walls of the channel. Preliminary results suggest that this effect makes the migration angle of the particles significantly different from that obtained by assuming a strictly two-dimensional structure for the flow of the suspending fluid.

scholarly journals Label-free enrichment of primary human skeletal progenitor cells using deterministic lateral displacement

Lab on a Chip ◽  
2019 ◽  
Vol 19 (3) ◽  
pp. 513-523 ◽  
Author(s):  
Miguel Xavier ◽  
Stefan H. Holm ◽  
Jason P. Beech ◽  
Daniel Spencer ◽  
Jonas O. Tegenfeldt ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Progenitor Cells ◽  
Cell Sorting ◽  
Lateral Displacement ◽  
Label Free ◽  
Deterministic Lateral Displacement ◽  
Cell Enrichment ◽  
Free Enrichment ◽  
Skeletal Stem Cell

Cell sorting using label-free microfluidic approaches offer significant promise for skeletal stem cell enrichment from human bone marrow.

Enhancing the Cell Viability in High Throughput Deterministic Lateral Displacement Separation of Circulating Tumor Cells

2019 ◽  
Author(s):  
Arian Aghilinejad ◽  
Christopher Landry ◽  
George Cha ◽  
Xiaolin Chen
Keyword(s):  
Reynolds Number ◽  
Tumor Cells ◽  
Circulating Tumor Cells ◽  
High Throughput ◽  
Cancer Metastasis ◽  
Lateral Displacement ◽  
Shear Stresses ◽  
Label Free ◽  
Deterministic Lateral Displacement ◽  
Reynolds Number Flow

Abstract Cancer is among a major health concerns all over the world. Cancer metastasis, which defines as the migration of malignant cells from original sites to distant organs, is the main reason of death due to cancer and there is growing evidence that Circulating Tumor Cells (CTCs) are responsible for initiating the metastasis. Due to the importance of these bioparticles in biotechnology and medicine, there is a growing interest to study and separate them through different techniques especially microfluidic label-free technologies. One such technology, termed Deterministic Lateral Displacement (DLD) has recently shown promising abilities to separate cells and particles of different sizes. However, DLD is a separation method that takes advantages of the predictable flow laminae of low Reynolds number (Re) fluid flow. In order to achieve higher device throughput, effects of higher Reynolds number flow on DLD device should be studied. Additionally, the higher flow rates would apply higher forces and shear stresses on the cells which threaten the cell’s viability. In this study, employing numerical simulation, the effect of high Re number on DLD device for separating cancer cells has been investigated. Specifically, we conducted force analysis and by focusing on the downstream gap distance between the posts, we improved the device which results in less cell deformation. Our developed numerical model and presented results lay the groundwork for design and fabrication of high-throughput DLD microchips for enhanced separation of CTCs.

scholarly journals Cell Sorting Using Electrokinetic Deterministic Lateral Displacement

Micromachines ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 30
Author(s):  
Bao D. Ho ◽  
Jason P. Beech ◽  
Jonas O. Tegenfeldt
Keyword(s):  
Zeta Potential ◽  
Lateral Displacement ◽  
Low Cost ◽  
Low Frequency ◽  
Cell Types ◽  
Considerable Change ◽  
Label Free ◽  
Size Change ◽  
Deterministic Lateral Displacement ◽  
Different Cell Types

We show that by combining deterministic lateral displacement (DLD) with electrokinetics, it is possible to sort cells based on differences in their membrane and/or internal structures. Using heat to deactivate cells, which change their viability and structure, we then demonstrate sorting of a mixture of viable and non-viable cells for two different cell types. For Escherichia coli, the size change due to deactivation is insufficient to allow size-based DLD separation. Our method instead leverages the considerable change in zeta potential to achieve separation at low frequency. Conversely, for Saccharomyces cerevisiae (Baker’s yeast) the heat treatment does not result in any significant change of zeta potential. Instead, we perform the sorting at higher frequency and utilize what we believe is a change in dielectrophoretic mobility for the separation. We expect our work to form a basis for the development of simple, low-cost, continuous label-free methods that can separate cells and bioparticles based on their intrinsic properties.

scholarly journals Label-Free Mass Spectrometry-Based Quantification of Linker Histone H1 Variants in Clinical Samples

2020 ◽  
Vol 21 (19) ◽  
pp. 7330
Author(s):  
Roberta Noberini ◽  
Cristina Morales Torres ◽  
Evelyn Oliva Savoia ◽  
Stefania Brandini ◽  
Maria Giovanna Jodice ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Histone H1 ◽  
Histone Variants ◽  
Linker Histone ◽  
Clinical Samples ◽  
Label Free ◽  
Complex Samples ◽  
Linker Histone H1 ◽  
Ideal Tool ◽  
Free Quantification

Epigenetic aberrations have been recognized as important contributors to cancer onset and development, and increasing evidence suggests that linker histone H1 variants may serve as biomarkers useful for patient stratification, as well as play an important role as drivers in cancer. Although traditionally histone H1 levels have been studied using antibody-based methods and RNA expression, these approaches suffer from limitations. Mass spectrometry (MS)-based proteomics represents the ideal tool to accurately quantify relative changes in protein abundance within complex samples. In this study, we used a label-free quantification approach to simultaneously analyze all somatic histone H1 variants in clinical samples and verified its applicability to laser micro-dissected tissue areas containing as low as 1000 cells. We then applied it to breast cancer patient samples, identifying differences in linker histone variants patters in primary triple-negative breast tumors with and without relapse after chemotherapy. This study highlights how label-free quantitation by MS is a valuable option to accurately quantitate histone H1 levels in different types of clinical samples, including very low-abundance patient tissues.

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