Sheathless High-Throughput Circulating Tumor Cell Separation Using Viscoelastic non-Newtonian Fluid

Circulating tumor cells (CTCs) have attracted increasing attention as important biomarkers for clinical and biological applications. Several microfluidic approaches have been demonstrated to separate CTCs using immunoaffinity or size difference from other blood cells. This study demonstrates a sheathless, high-throughput separation of CTCs from white blood cells (WBCs) using a viscoelastic fluid. To determine the fluid viscoelasticity and the flow rate for CTC separation, and to validate the device performance, flow characteristics of 6, 13, and 27 μm particles in viscoelastic fluids with various concentrations were estimated at different flow rates. Using 0.2% hyaluronic acid (HA) solution, MCF-7 (Michigan Cancer Foundation-7) cells mimicking CTCs in this study were successfully separated from WBCs at 500 μL/min with a separation efficiency of 94.8%. Small amounts of MCF-7 cells (~5.2%) were found at the center outlet due to the size overlap with WBCs.

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High-throughput white blood cells (leukocytes) separation and enrichment from whole blood by hydrodynamic and inertial force

2012 IEEE 25th International Conference on Micro Electro Mechanical Systems (MEMS) ◽

10.1109/memsys.2012.6170315 ◽

2012 ◽

Cited By ~ 2

Author(s):

Horas-Cendana Tseng ◽

Ren-Guei Wu ◽

Hwan-You Chang ◽

Fan-Gang Tseng

Keyword(s):

High Throughput ◽

Whole Blood ◽

Blood Cells ◽

White Blood Cells ◽

Inertial Force

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Design and Simulation of an Integrated Centrifugal Microfluidic Device for CTCs Separation and Cell Lysis

Micromachines ◽

10.3390/mi11070699 ◽

2020 ◽

Vol 11 (7) ◽

pp. 699

Author(s):

Rohollah Nasiri ◽

Amir Shamloo ◽

Javad Akbari ◽

Peyton Tebon ◽

Mehmet R. Dokmeci ◽

...

Keyword(s):

Numerical Simulation ◽

Angular Velocity ◽

Microfluidic Device ◽

Cell Separation ◽

High Efficiency ◽

Separation Efficiency ◽

White Blood Cells ◽

Cell Lysis ◽

Target Cells ◽

Separation Unit

Separation of circulating tumor cells (CTCs) from blood samples and subsequent DNA extraction from these cells play a crucial role in cancer research and drug discovery. Microfluidics is a versatile technology that has been applied to create niche solutions to biomedical applications, such as cell separation and mixing, droplet generation, bioprinting, and organs on a chip. Centrifugal microfluidic biochips created on compact disks show great potential in processing biological samples for point of care diagnostics. This study investigates the design and numerical simulation of an integrated microfluidic device, including a cell separation unit for isolating CTCs from a blood sample and a micromixer unit for cell lysis on a rotating disk platform. For this purpose, an inertial microfluidic device was designed for the separation of target cells by using contraction–expansion microchannel arrays. Additionally, a micromixer was incorporated to mix separated target cells with the cell lysis chemical reagent to dissolve their membranes to facilitate further assays. Our numerical simulation approach was validated for both cell separation and micromixer units and corroborates existing experimental results. In the first compartment of the proposed device (cell separation unit), several simulations were performed at different angular velocities from 500 rpm to 3000 rpm to find the optimum angular velocity for maximum separation efficiency. By using the proposed inertial separation approach, CTCs, were successfully separated from white blood cells (WBCs) with high efficiency (~90%) at an angular velocity of 2000 rpm. Furthermore, a serpentine channel with rectangular obstacles was designed to achieve a highly efficient micromixer unit with high mixing quality (~98%) for isolated CTCs lysis at 2000 rpm.

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Acoustic Cell Separation Based on Density and Mechanical Properties

Journal of Biomechanical Engineering ◽

10.1115/1.4046180 ◽

2020 ◽

Vol 142 (3) ◽

Cited By ~ 4

Author(s):

Yuliang Xie ◽

Zhangming Mao ◽

Hunter Bachman ◽

Peng Li ◽

Peiran Zhang ◽

...

Keyword(s):

Mechanical Properties ◽

Blood Cells ◽

Recovery Rate ◽

Cell Separation ◽

White Blood Cells ◽

Acoustic Properties ◽

Biological Research ◽

Microfluidic Channels ◽

Fluid Media ◽

Solution Surface

Abstract Density and mechanical properties (e.g., compressibility or bulk modulus) are important cellular biophysical markers. As such, developing a method to separate cells directly based on these properties can benefit various applications including biological research, diagnosis, prognosis, and therapeutics. As a potential solution, surface acoustic wave (SAW)-based cell separation has demonstrated advantages in terms of biocompatibility and compact device size. However, most SAW-reliant cell separations are achieved using an entangled effect of density, various mechanical properties, and size. In this work, we demonstrate SAW-based separation of cells/particles based on their density and compressibility, irrespective of their sizes, by manipulating the acoustic properties of the fluidic medium. Using our platform, SAW-based separation is achieved by varying the dimensions of the microfluidic channels, the wavelengths of acoustic signals, and the properties of the fluid media. Our method was applied to separate paraformaldehyde-treated and fresh Hela cells based on differences in mechanical properties; a recovery rate of 85% for fixed cells was achieved. It was also applied to separate red blood cells (RBCs) and white blood cells (WBCs) which have different densities. A recovery rate of 80.5% for WBCs was achieved.

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Discrimination between Breast Cancer Cells and White Blood Cells by Non-Invasive Measurements: Implications for a Novel In Vitro-Based Circulating Tumor Cell Model Using Digital Holographic Cytometry

Applied Sciences ◽

10.3390/app10144854 ◽

2020 ◽

Vol 10 (14) ◽

pp. 4854

Author(s):

Zahra El-Schich ◽

Birgit Janicke ◽

Kersti Alm ◽

Nishtman Dizeyi ◽

Jenny L. Persson ◽

...

Keyword(s):

Breast Cancer ◽

Tumor Cell ◽

Cancer Cells ◽

Cell Lines ◽

Blood Cells ◽

Breast Cancer Cells ◽

Cell Model ◽

White Blood Cells ◽

Circulating Tumor Cell

Breast cancer is the second most common cancer worldwide. Metastasis is the main reason for death in breast cancer, and today, there is a lack of methods to detect and isolate circulating tumor cells (CTCs), mainly due to their heterogeneity and rarity. There are some systems that are designed to detect rare epithelial cancer cells in whole blood based on the most common marker used today, the epithelial cell adhesion molecule (EpCAM). It has been shown that aggressive breast cancer metastases are of non-epithelial origin and are therefore not always detected using EpCAM as a marker. In the present study, we used an in vitro-based circulating tumor cell model comprising a collection of six breast cancer cell lines and white blood cell lines. We used digital holographic cytometry (DHC) to characterize and distinguish between the different cell types by area, volume and thickness. Here, we present significant differences in cell size-related parameters observed when comparing white blood cells and breast cancer cells by using DHC. In conclusion, DHC can be a powerful diagnostic tool for the characterization of CTCs in the blood.

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Detection of live breast cancer cells in brightfield microscopy images containing white blood cells by image analysis and deep learning

10.1101/2021.11.04.467215 ◽

2021 ◽

Author(s):

Golnaz Moallem ◽

Adity A. Pore ◽

Anirudh Gangadhar ◽

Hamed Sari-Sarraf ◽

Siva A Vanapalli

Keyword(s):

Breast Cancer ◽

Neural Network ◽

Deep Learning ◽

Convolutional Neural Network ◽

Cancer Cells ◽

Blood Cells ◽

Breast Cancer Cells ◽

White Blood Cells ◽

Size Difference ◽

Microscopy Images

Significance: Circulating tumor cells (CTCs) are important biomarkers for cancer management. Isolated CTCs from blood are stained to detect and enumerate CTCs. However, the staining process is laborious and moreover makes CTCs unsuitable for drug testing and molecular characterization. Aim: The goal is to develop and test deep learning (DL) approaches to detect unstained breast cancer cells in bright field microscopy images that contain white blood cells (WBCs). Approach: We tested two convolutional neural network (CNN) approaches. The first approach allows investigation of the prominent features extracted by CNN to discriminate cancer cells from WBCs. The second approach is based on Faster Region-based Convolutional Neural Network (Faster R-CNN). Results: Both approaches detected cancer cells with high sensitivity and specificity with the Faster R-CNN being more efficient and suitable for deployment. The distinctive feature used by the CNN used to discriminate is cell size, however, in the absence of size difference, the CNN was found to be capable of learning other features. The Faster R-CNN was found to be robust with respect to intensity and contrast image transformations. Conclusions: CNN-based deep learning approaches could be potentially applied to detect patient-derived CTCs from images of blood samples.

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High-Throughput, Label-Free Isolation of White Blood Cells From Whole Blood Using Parallel Spiral Microchannels With U-Shaped Cross-Section

10.20944/preprints202109.0482.v1 ◽

2021 ◽

Author(s):

Amirhossein Mehran ◽

Peyman Rostami ◽

Mohammad Said Saidi ◽

Bahar Firoozabadi ◽

Navid Kashaninejad

Keyword(s):

Cross Section ◽

High Throughput ◽

Whole Blood ◽

Blood Cells ◽

High Efficiency ◽

Rectangular Cross Section ◽

White Blood Cells ◽

Label Free ◽

Entire Cross Section ◽

Shaped Cross Section

Rapid isolation of white blood cells (WBCs) from whole blood is an essential part of any WBC examination platform. However, most conventional cell separation techniques are labor-intensive and low throughput, require large volumes of samples, need extensive cell manipulation, and have low purity. To address these challenges, we report the design and fabrication of a passive, label-free microfluidic device with a unique U-shaped cross-section to separate WBCs from whole blood using hydrodynamic forces that exist in a microchannel with curvilinear geometry. It is shown that the spiral microchannel with a U-shaped cross-section concentrates larger blood cells (e.g., WBCs) in the inner cross-section of the microchannel by moving smaller blood cells (e.g., red blood cells (RBCs) and platelets) to the outer microchannel section and preventing them from returning to the inner microchannel section. Therefore, it overcomes the major limitation of a rectangular cross-section where secondary Dean vortices constantly enforce particles throughout the entire cross-section and decrease its isolation efficiency. Under optimal settings, more than 95% of WBCs can be isolated from whole blood under high-throughput (6 ml/min), high-purity (88%), and high-capacity (180 ml of sample in 1 hour) conditions. High efficiency, fast processing time, and non-invasive WBC isolation from large blood samples without centrifugation, RBC lysis, cell biomarkers, and chemical pre-treatments make this method an ideal choice for downstream cell study platforms.

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