Size-based hydrodynamic rare tumor cell separation in curved microfluidic channels.
Ontology highlight
ABSTRACT:
SUBMITTER: Sun J
PROVIDER: S-EPMC3555910 | biostudies-other | 2013
REPOSITORIES: biostudies-other
Similar Datasets
Project description:The present work demonstrates the use of a dielectrophoretic lab-on-a-chip device in effectively separating different cancer cells of epithelial origin for application in circulating tumor cell (CTC) identification. This study uses dielectrophoresis (DEP) to distinguish and separate MCF-7 human breast cancer cells from HCT-116 colorectal cancer cells. The DEP responses for each cell type were measured against AC electrical frequency changes in solutions of varying conductivities. Increasing the conductivity of the suspension directly correlated with an increasing frequency value for the first cross-over (no DEP force) point in the DEP spectra. Differences in the cross-over frequency for each cell type were leveraged to determine a frequency at which the two types of cell could be separated through DEP forces. Under a particular medium conductivity, different types of cells could have different DEP behaviors in a very narrow AC frequency band, demonstrating a high specificity of DEP. Using a microfluidic DEP sorter with optically transparent electrodes, MCF-7 and HCT-116 cells were successfully separated from each other under a 3.2 MHz frequency in a 0.1X PBS solution. Further experiments were conducted to characterize the separation efficiency (enrichment factor) by changing experimental parameters (AC frequency, voltage, and flow rate). This work has shown the high specificity of the described DEP cell sorter for distinguishing cells with similar characteristics for potential diagnostic applications through CTC enrichment.
| S-EPMC3555970 | biostudies-other
Project description:Blood cells naturally auto-segregate in postcapillary venules, with the erythrocytes (red blood cells, RBCs) aggregating near the axis of flow and the nucleated cells (NCs)--which include leukocytes, progenitor cells and, in cancer patients, circulating tumor cells--marginating toward the vessel wall. We have used this principle to design a microfluidic device that extracts nucleated cells (NCs) from whole blood. Fabricated using polydimethylsiloxane (PDMS) soft lithography, the biomimetic cell extraction device consists of rectangular microchannels that are 20-400 ?m wide, 11 ?m deep and up to 2 cm long. The key design feature is the use of repeated expansions/contractions of triangular geometry mimicking postcapillary venules, which enhance margination and optimize the extraction. The device operates on unprocessed whole blood and is able to extract 94 ± 4.5% of NCs with 45.75 ± 2.5-fold enrichment in concentration at a rate of 5 nl s(-1). The device eliminates the need to preprocess blood via centrifugation or RBC lysis, and is ready to be implemented as the initial stage of lab-on-a-chip devices that require enriched nucleated cells. The potential downstream applications are numerous, encompassing all preclinical and clinical assays that operate on enriched NC populations and include on-chip flow cytometry (A. Y. Fu et al., Anal. Chem., 2002, 74, 2451-2457; A. Y. Fu et al., Nat. Biotechnol., 1999, 17, 1109-1111), genetic analyses (M. M. Wang et al., Nat. Biotechnol., 2005, 23, 83-87; L. C. Waters et al., Anal. Chem., 1998, 70, 5172-5176) and circulating tumor cell extraction (S. Nagrath et al., Nature, 2007, 450, 1235-1241; S. L. Stott et al., Proc. Natl. Acad. Sci. U. S. A., 2010, 18392-18397; H. K. Lin et al., Clin. Cancer Res., 2010, 16, 5011-5018).
| S-EPMC3743538 | biostudies-literature
Project description:Circulating tumor cells (CTCs) were introduced as biomarkers more than 10 years ago, but capture of viable CTCs at high purity from peripheral blood of cancer patients is still a major technical challenge. Here, we report a novel microfluidic platform designed for marker independent capture of CTCs. The Parsortix™ cell separation system provides size and deformability-based enrichment with automated staining for cell identification, and subsequent recovery (harvesting) of cells from the device. Using the Parsortix™ system, average cell capture inside the device ranged between 42% and 70%. Subsequent harvest of cells from the device ranged between 54% and 69% of cells captured. Most importantly, 99% of the isolated tumor cells were viable after processing in spiking experiments as well as after harvesting from patient samples and still functional for downstream molecular analysis as demonstrated by mRNA characterization and array-based comparative genomic hybridization. Analyzing clinical blood samples from metastatic (n = 20) and nonmetastatic (n = 6) cancer patients in parallel with CellSearch(®) system, we found that there was no statistically significant difference between the quantitative behavior of the two systems in this set of twenty six paired separations. In conclusion, the epitope independent Parsortix™ system enables the isolation of viable CTCs at a very high purity. Using this system, viable tumor cells are easily accessible and ready for molecular and functional analysis. The system's ability for enumeration and molecular characterization of EpCAM-negative CTCs will help to broaden research into the mechanisms of cancer as well as facilitating the use of CTCs as "liquid biopsies."
| S-EPMC5069649 | biostudies-literature
Project description:With a growing number of cell-based biotechnological applications, there is a need for particle separation systems capable of multiparameter separations at high purity and throughput, beyond what is presently offered by traditional methods including fluorescence activated cell sorting and column-based magnetic separation. Toward this aim, we report on the integration of microfluidic acoustic and magnetic separation in a monolithic device for multiparameter particle separation. Using our device, we demonstrate high-purity separation of a multicomponent particle mixture at a throughput of up to 10(8) particleshr.
| S-EPMC2807449 | biostudies-other
Project description:Circulating tumor cell clusters (CTC clusters) are potent initiators of metastasis and potentially useful clinical markers for patients with cancer. Although there are numerous devices developed to isolate individual circulating tumor cells from blood, these devices are ineffective at capturing CTC clusters, incapable of separating clusters from single cells and/or cause cluster damage or dissociation during processing. The only device currently able to specifically isolate CTC clusters from single CTCs and blood cells relies on the batch immobilization of clusters onto micropillars which necessitates long residence times and causes damage to clusters during release. Here, we present a two-stage continuous microfluidic chip that isolates and recovers viable CTC clusters from blood. This approach uses deterministic lateral displacement to sort clusters by capitalizing on two geometric properties: size and asymmetry. Cultured breast cancer CTC clusters containing between 2-100 + cells were recovered from whole blood using this integrated two-stage device with minimal cluster dissociation, 99% recovery of large clusters, cell viabilities over 87% and greater than five-log depletion of red blood cells. This continuous-flow cluster chip will enable further studies examining CTC clusters in research and clinical applications.
| S-EPMC5446400 | biostudies-literature
Project description:Circulating tumor cells (CTCs) are extremely rare in the blood, yet they account for metastasis. Notably, it was reported that CTC clusters (CTCCs) can be 50-100 times more metastatic than single CTCs, making them particularly salient as a liquid biopsy target. Yet they can split apart and are even rarer, complicating their recovery. Isolation by filtration risks loss when clusters squeeze through filter pores over time, and release of captured clusters can be difficult. Deterministic lateral displacement is continuous but requires channels not much larger than clusters, leading to clogging. Spiral inertial focusing requires large blood dilution factors (or lysis). Here, we report a microfluidic chip that continuously isolates untouched CTC clusters from large volumes of minimally (or undiluted) whole blood. An array of 100 ?m-wide channels first concentrates clusters in the blood, and then a similar array transfers them into a small volume of buffer. The microscope-slide-sized PDMS device isolates individually-spiked CTC clusters from >30 mL per hour of whole blood with 80% efficiency into enumeration (fluorescence imaging), and on-chip yield approaches 100% (high speed video). Median blood cell removal (in base-10 logs) is 4.2 for leukocytes, 5.5 for red blood cells, and 4.9 for platelets, leaving less than 0.01% of leukocytes alongside CTC clusters in the product. We also demonstrate that cluster configurations are preserved. Gentle, high throughput concentration and separation of circulating tumor cell clusters from large blood volumes will enable cluster-specific diagnostics and speed the generation of patient-specific CTC cluster lines.
| S-EPMC7469923 | biostudies-literature
Project description:Particle separation techniques play an important role in biomedical research. Inertial focusing based microfluidics using nonlinear channels is one of the promising label-free technologies for biological applications. The particle separation is achieved as a result of the combination of inertial lift force (FL) and Dean drag force (FD). Although the mathematical expressions of FL and FD have been well derived in prior studies, they are still complicated, which limits their popularity in practice. Recent studies modified these expressions through experiments and proposed a threshold model, which assumes that only particles larger than the threshold will be well focused. Although this threshold model has been used in recent studies, two varying versions of the threshold model (TM1 and TM2) prevents standardisation in practice. In addition, both models were developed with regular low-density particles and may not be applicable to samples with higher density or samples with irregular shapes. Here, we evaluated the threshold models with samples of different densities. Based on these evaluations, we derived a modified model (TM4), which additionally considers the factor of particle density to improve the accuracy of existing models. Our results demonstrated that TM4 could more reliably predict the sorting efficiency of samples within a wider density range.
| S-EPMC7643107 | biostudies-literature
Project description:Microfluidic-based separation methods have been highlighted for a number of biological applications, such as single cell analysis, disease diagnostics, and therapeutics. Although a number of previous studies have been carried out to minimize the physical damage and chemical deformations of the sample during the separation process, it still remains a challenge. In this paper, we developed a microfluidic device with dual-neodymium magnet-based negative magnetophoresis for the separation of the microparticles and cells. The poly(ethylene oxide) (PEO) was added to the solution to increase the viscoelasticity of the medium which could assist the sorting of the microparticles in the microfluidic device even at low flow rates, while minimizing damage to the cells and microparticles. Following this method, it was possible to separate 10 and 16 μm microparticles with high efficiency of 99 ± 0.1%, and 97 ± 0.8%, respectively. We also demonstrated the separation of glioblastoma cancer cells and neural stem cells (NSCs) in the microfluidic device.
| S-EPMC6602928 | biostudies-literature
Project description:Two size-based separation strategies are evaluated for suspensions consisting of giant unilamellar vesicles with a broad, continuous distribution of diameters. Microfluidic devices were designed to separate an initial suspension into larger and smaller particles via either filtration or inertial focusing. These separation mechanisms were tested with suspensions of vesicles and suspensions of rigid spheres separately to illustrate the effect of deformability on separation ability. We define several separation metrics to assess the separation ability and to enable comparison between separation strategies. The filtration device significantly reduced the polydispersity of the separated vesicle fractions relative to the starting suspension and displayed an ability to separate vesicle suspensions at high throughputs. The device that utilized inertial focusing exhibited adequate polydispersity reduction and performed best with diluted vesicle suspensions. The inertial device had fewer issues with debris and trapped air, leading to short device preparation times and indicating a potential for continuous separation operation.
| S-EPMC5446298 | biostudies-literature
Project description:Cell isolation in designated regions or from heterogeneous samples is often required for many microfluidic cell-based assays. However, current techniques have either limited throughput or are incapable of viable off-chip collection. We present an innovative approach, allowing high-throughput and label-free cell isolation and enrichment from heterogeneous solution using cell size as a biomarker. The approach utilizes the irreversible migration of particles into microscale vortices, developed in parallel expansion-contraction trapping reservoirs, as the cell isolation mechanism. We empirically determined the critical particle∕cell diameter D(crt) and the operational flow rate above which trapping of cells∕particles in microvortices is initiated. Using this approach we successfully separated larger cancer cells spiked in blood from the smaller blood cells with processing rates as high as 7.5×10(6) cells∕s. Viable long-term culture was established using cells collected off-chip, suggesting that the proposed technique would be useful for clinical and research applications in which in vitro culture is often desired. The presented technology improves on current technology by enriching cells based on size without clogging mechanical filters, employing only a simple single-layered microfluidic device and processing cell solutions at the ml∕min scale.
| S-EPMC3171489 | biostudies-other