title: Label-free impedance detection of cancer cells from whole blood on an integrated centrifugal microfluidic platform.
abstract: An electrochemical Lab-on-a-Disc (eLoaD) platform for the automated quantification of ovarian cancer cells (SKOV3) from whole blood is reported. This centrifugal microfluidic system combines complex sample handling, i.e., blood separation and cancer cell extraction from plasma, with specific capture and sensitive detection using label-free electrochemical impedance. Flow control is facilitated using rotationally actuated valving strategies including siphoning, capillary and centrifugo-pneumatic dissolvable-film (DF) valves. For the detection systems, the thiol-containing amino acid, L-Cysteine, was self-assembled onto smooth gold electrodes and functionalized with anti-EpCAM. By adjusting the concentration of buffer electrolyte, the thickness of the electrical double layer was extended so the interfacial electric field interacts with the bound cells. Significant impedance changes were recorded at 117.2 Hz and 46.5 Hz upon cell capture. Applying AC amplitude of 50 mV at 117.2 Hz and open circuit potential, a minimum of 214 captured cells/mm(2) and 87% capture efficiency could be recorded. The eLoaD platform can perform five different assays in parallel with linear dynamic range between 16,400 and (2.6±0.0003)×10(6) cancer cells/mL of blood, i.e. covering nearly three orders of magnitude. Using the electrode area of 15.3 mm(2) and an SKOV3 cell radius of 5 µm, the lower detection limit is equivalent to a fractional surface coverage of approximately 2%, thus making eLoaD a highly sensitive and efficient prognostic tool that can be developed for clinical settings where ease of handling and minimal sample preparation are paramount.
authors.affiliation: Biomedical Diagnostics Institute, National Center for Sensor Research, Dublin City University, Ireland; School of Physical Sciences, Dublin City University, Ireland. Electronic address: charles.nwankire@dcu.ie.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Nwankire
authors.affiliation: Biomedical Diagnostics Institute, National Center for Sensor Research, Dublin City University, Ireland; School of Chemical Sciences, Dublin City University, Ireland.
authors.forename: Anita
authors.initials: A
authors.lastname: Venkatanarayanan
authors.affiliation: Biomedical Diagnostics Institute, National Center for Sensor Research, Dublin City University, Ireland; School of Physical Sciences, Dublin City University, Ireland.
authors.forename: Thomas
authors.initials: T
authors.lastname: Glennon
authors.affiliation: Biomedical Diagnostics Institute, National Center for Sensor Research, Dublin City University, Ireland; School of Chemical Sciences, Dublin City University, Ireland.
authors.forename: Tia E
authors.initials: TE
authors.lastname: Keyes
authors.affiliation: Biomedical Diagnostics Institute, National Center for Sensor Research, Dublin City University, Ireland; School of Chemical Sciences, Dublin City University, Ireland.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Forster
authors.affiliation: Biomedical Diagnostics Institute, National Center for Sensor Research, Dublin City University, Ireland; School of Physical Sciences, Dublin City University, Ireland. Electronic address: jens.ducree@dcu.ie.
authors.forename: Jens
authors.initials: J
authors.lastname: Ducrée
mesh_terms: D055910:Antibodies, Immobilized / Q000276:immunology; D015374:Biosensing Techniques / Q000295:instrumentation*; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000295:instrumentation; D002498:Centrifugation / Q000295:instrumentation; D058266:Dielectric Spectroscopy / Q000295:instrumentation*; D004867:Equipment Design; D005260:Female; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation; D057230:Limit of Detection; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009360:Neoplastic Cells, Circulating / Q000276:immunology / Q000473:pathology*; D010051:Ovarian Neoplasms / Q000097:blood* / Q000276:immunology / Q000473:pathology*; D010053:Ovary / Q000276:immunology / Q000473:pathology
chemical_list: D055910:Antibodies, Immobilized
keywords: Biosensor; Cancer; Centrifugal microfluidics; Circulating tumour cells; Electrochemical; EpCAM; Immunoassay; Impedance; Lab-on-a-Disc; Label-free detection