title: In situ single cell detection via microfluidic magnetic bead assay.
abstract: We present a single cell detection device based on magnetic bead assay and micro Coulter counters. This device consists of two successive micro Coulter counters, coupled with a high gradient magnetic field generated by an external magnet. The device can identify single cells in terms of the transit time difference of the cell through the two micro Coulter counters. Target cells are conjugated with magnetic beads via specific antibody and antigen binding. A target cell traveling through the two Coulter counters interacts with the magnetic field, and have a longer transit time at the 1st counter than that at the 2nd counter. In comparison, a non-target cell has no interaction with the magnetic field, and hence has nearly the same transit times through the two counters. Each cell passing through the two counters generates two consecutive voltage pulses one after the other; the pulse widths and magnitudes indicating the cell's transit times through the counters and the cell's size respectively. Thus, by measuring the pulse widths (transit times) of each cell through the two counters, each single target cell can be differentiated from non-target cells even if they have similar sizes. We experimentally proved that the target human umbilical vein endothelial cells (HUVECs) and non-target rat adipose-derived stem cells (rASCs) have significant different transit time distribution, from which we can determine the recognition regions for both cell groups quantitatively. We further demonstrated that within a mixed cell population of rASCs and HUVECs, HUVECs can be detected in situ and the measured HUVECs ratios agree well with the pre-set ratios. With the simple device structure and easy sample preparation, this method is expected to enable single cell detection in a continuous flow and can be applied to facilitate general cell detection applications such as stem cell identification and enumeration.
authors.affiliation: Department of Mechanical Engineering, University of Akron, Akron, Ohio, United States of America.
authors.forename: Fan
authors.identifier: http://orcid.org/0000-0001-8827-2278
authors.initials: F
authors.lastname: Liu
authors.affiliation: Department of Biomedical Engineering, University of Akron, Akron, Ohio, United States of America.
authors.forename: Pawan
authors.initials: P
authors.lastname: Kc
authors.affiliation: Department of Biomedical Engineering, University of Akron, Akron, Ohio, United States of America.
authors.forename: Ge
authors.initials: G
authors.lastname: Zhang
authors.affiliation: Department of Mechanical Engineering, University of Akron, Akron, Ohio, United States of America.
authors.forename: Jiang
authors.initials: J
authors.lastname: Zhe
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D002138:Calibration; D002452:Cell Count; D002460:Cell Line; D048429:Cell Size; D004867:Equipment Design; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D018189:Immunomagnetic Separation / Q000295:instrumentation*; D056656:Lab-On-A-Chip Devices; D046210:Microfluidic Analytical Techniques; D008863:Microspheres; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000276:immunology; D051381:Rats; D059010:Single-Cell Analysis / Q000295:instrumentation / Q000379:methods*; D013045:Species Specificity; D013234:Stem Cells
chemical_list: D019408:Platelet Endothelial Cell Adhesion Molecule-1