Magnetophoretic Cell Sorting Is a Function of Antibody Binding Capacity
61
Citation
0
Reference
10
Related Paper
Citation Trend
Abstract:
Antibody binding capacity (ABC) is a term representing a cellapos;s ability to bind antibodies, correlating with the number of specific cellular antigens expressed on that cell. ABC allows magnetically conjugated antibodies to bind to the targeted cells, imparting a magnetophoretic mobility on each targeted cell. This enables sorting based on differences in the cell magnetophoretic mobility and, potentially, a magnetic separation based on the differences in the cell ABC values. A cellapos;s ABC value is a particularly important factor in continuous magnetic cell separation. This work investigates the relationship between ABC and magnetic cell separation efficiency by injection of a suspension of immunomagnetically labeled quantum simply cellular calibration microbeads of known ABC values into fluid flowing through a quadrupole magnetic sorter. The elution profiles of the outlet streams were evaluated using UV detectors. Optimal separation flow rate was shown to correlate with the ABC of these microbeads. Comparing experimental and theoretical results, the theory correctly predicted maximum separation flow rates but overestimated the separation fractional recoveries.Keywords:
Cell Sorting
Magnetic separation
Immunomagnetic separation
Circulating tumor cell
Immunomagnetic separation
Cell Sorting
Cite
Citations (41)
Immunomagnetic separation
Cell Sorting
Cell type
Cite
Citations (22)
Cell Sorting
Cytometry
Cite
Citations (135)
Micro total analysis systems (µTAS) also referred to as "lab-on-a-chip" is one of the fastest progressing fields in biological and chemical analyses. In recent years, µTAS for single cell analysis has drawn the attention of researchers due to its significant advantages over traditional methods for single cell manipulation, fast cell sorting and integration of multiple functions. As the preliminary step for studying cells on chips, cell sorting using microfluidics have been investigated by researchers intensively. This article reviews the most recent advances on microfuidics-based cell sorting techniques including cell sorting principle, strategy, mechanism and procedure with emphases on the sorting mechanism and procedure. Furthermore, evaluation criteria for successful cell sorter are also discussed and future research directions are given.
Cell Sorting
Microfluidic chip
Cite
Citations (58)
Enrichment and purification of bacteria from complex matrices are crucial for their detection and investigation, in which magnetic separation techniques have recently show great application advantages. However, currently used magnetic particles all have their own limitations: Magnetic microparticles exhibit poor binding capacity with targets, while magnetic nanoparticles suffer slow magnetic response and high loss rate during treatment process. Herein, we used a highly controllable layer-by-layer assembly method to fabricate quick-response magnetic nanospheres (MNs), and with Salmonella typhimurium as a model, we successfully achieve their rapid and efficient enrichment. The MNs combined the advantages of magnetic microparticles and nanoparticles. On the one hand, the MNs had a fast magnetic response, and almost 100% of the MNs could be recovered by 1 min attraction with a simple magnetic scaffold. Hence, using antibody conjugated MNs (immunomagnetic nanospheres, IMNs) to capture bacteria hardly generated loss and did not need complex separation tools or techniques. On the other hand, the IMNs showed much excellent capture capacity. With 20 min interaction, almost all of the target bacteria could be captured, and even only one bacterium existing in the samples was not missed, comparing with the immunomagnetic microparticles which could only capture less than 50% of the bacteria. Besides, the IMNs could achieve the same efficient enrichment in complex matrices, such as milk, fetal bovine serum, and urine, demonstrating their good stability, strong anti-interference ability, and low nonspecific adsorption. In addition, the isolated bacteria could be directly used for culture, polymerase chain reaction (PCR) analyses, and fluorescence immunoassay without a release process, which suggested our IMNs-based enrichment strategy could be conveniently coupled with the downstream identification and analysis techniques. Thus, the MNs provided by this work showed great superiority in bacteria enrichment, which would be a promising tool for bacteria detection and investigation.
Magnetic separation
Immunomagnetic separation
Cite
Citations (51)
We present a new approach for sorting cells based upon visual information, termed opto-fluidic cell sorting (OPTO-FluCS). This technique uses a combination of hydrodynamic and optical scattering forces to array, image, and then sort many individual cells. Our device combines the strengths of microscopy–imaging in space and time–with the ability to array cell populations and isolate cells of interest in an intuitive, user-friendly fashion. We successfully demonstrate image-based sorting based on fluorescence localization, a sort unavailable with flow-assisted cell sorting.
Cell Sorting
Fluidics
Cite
Citations (1)
Recent researches on single cell analysis have provided important results showing differences between the individual cells even in the same culture group. On-chip cell sorting techniques represented by the fluorescent-activated cell sorting (FACS) using microfluidic chips have greatly contributed to these analyses because of their remarkable high-throughput ability. Currently, the cell sorting faces to a chance of further leap with the aid of microfabrication technologies and microfluidic flow control technologies for constructing the high-throughput system. In this article, we give a commentary for the on-chip cell sorting from the sight of sorting methods.
Cell Sorting
Microfluidic chip
Lab-on-a-Chip
Cite
Citations (0)
Abstract Processing of peripheral blood progenitor cells (PBPC) for clinical transplantation or research applications aims to effectively isolate or deplete specific cell populations, utilizing primarily magnetic or fluorescence activated sorting methods. Here, we investigated the performance of microfluidic acoustophoresis for the separation of lymphocyte subsets from PBPC, and present a novel method for affinity‐bead‐mediated acoustic separation of cells which can otherwise not be acoustically discriminated. As the acoustic force on a particle depends on particle size, density and compressibility, targeting of cells by affinity specific beads will generate cell‐bead complexes that exhibit distinct acoustic properties relative to nontargeted cells and are, thus, possible to isolate. To demonstrate this, PBPC samples ( n = 22) were obtained from patients and healthy donors. Following density gradient centrifugation, cells were labeled with anti‐CD4‐coated magnetic beads (Dynal) and isolated by acoustophoresis and, for comparison, standard magnetic cell sorting technique in parallel. Targeted CD4+ lymphocytes were acoustically isolated with a mean (±SD) purity of 87 ± 12%, compared with 96 ± 3% for control magnetic sorting. Viability of sorted cells was 95 ± 4% (acoustic) and 97 ± 3% (magnetic), respectively. The mean acoustic separation efficiency of CD4+ lymphocytes to the target fraction was 65 ± 22%, compared with a mean CD4+ lymphocyte recovery of 56 ± 15% for magnetic sorting. Functional testing of targeted CD4+ lymphocytes demonstrated unimpaired mitogen‐mediated proliferation capacity and cytokine production. Hematopoietic progenitor cell assays revealed a preserved colony forming ability of nontarget cells post sorting. We conclude that the acoustophoresis platform can be utilized to efficiently isolate bead‐labeled CD4+ lymphocytes from PBPC samples in a continuous flow format, with preserved functional capacity of both target and nontarget cells. These results open up for simultaneous affinity‐bead‐mediated separation of multiple cell populations, something which is not possible with current standard magnetic cell separation technology. © 2014 International Society for Advancement of Cytometry
Cell Sorting
Immunomagnetic separation
Differential centrifugation
Magnetic separation
Magnetic bead
Bead
Cite
Citations (34)
We present a microfluidic cell-sorting device which augments microscopy with the capability to perform facile image-based cell sorting. This combination enables intuitive, complex phenotype sorting based on spatio−temporal fluorescence or cell morphology. The microfluidic device contains a microwell array that can be passively loaded with mammalian cells via sedimentation and can be subsequently inspected with microscopy. After inspection, we use the scattering force from a focused infrared laser to levitate cells of interest from their wells into a flow field for collection. First, we demonstrate image-based sorting predicated on whole-cell fluorescence, which could enable sorting based on temporal whole-cell fluorescence behavior. Second, we demonstrate image-based sorting predicated on fluorescence localization (nuclear vs whole-cell fluorescence), highlighting the capability of our approach to sort based on imaged subcellular events, such as localized protein expression or translocation events. We achieve postsort purities up to 89% and up to 155-fold enrichment of target cells. Optical manipulation literature and a direct cell viability assay suggest that cells remain viable after using our technique. The architecture is highly scalable and supports over 10 000 individually addressable trap sites. Our approach enables sorting of significant populations based on subcellular spatio−temporal information, which is difficult or impossible with existing widespread sorting technologies.
Cell Sorting
Cite
Citations (121)
A novel platform, called SeParate, enabling accurate multiplex droplet sorting by integrating serial and parallel sorting principles for three model systems with increasing complexity and intra-subpopulation variation in fluorescence intensities.
Cite
Citations (4)