title: Label-free method for cell counting in crude biological samples via paramagnetic bead aggregation.
abstract: Under chaotropic conditions, DNA released from lysed cells causes the aggregation of paramagnetic beads in a rotating magnetic field in a manner that is independent of the presence of other cellular components. The extent of aggregation correlates with the mass of DNA in a quantitative manner (Leslie, D. C. et al., J. Am. Chem. Soc. 2012, 134, 5689-96), and from this, the number of DNA-containing cells in the sample can be enumerated. Microbial growth testing is demonstrated by monitoring bead aggregation with E. coli in the presence of ampicillin. Without the need for fluorescent labeling or Coulter counting, the white blood cell count can be defined directly from a microliter of crude whole blood. Specificity is brought to the process by coupling bead-based immunocapture with DNA-bead aggregation allowing for the enumeration of CD4+ T cells from human blood samples. The results of DNA-induced bead aggregation had a 95% correlation with those generated by flow cytometry. With the process requiring only inexpensive, widely available benchtop laboratory hardware, a digital camera, and a simple algorithm, this provided a highly accessible alternative to more expensive cell-counting techniques.
authors.affiliation: Department of Chemistry, ‡Department of Pathology, §Department of Biomedical Engineering, ⊥Department of Mechanical and Aerospace Engineering, ∥Center for Microsystems for the Life Sciences, University of Virginia , Charlottesville, VA 22904.
authors.forename: Jingyi
authors.initials: J
authors.lastname: Li
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.forename: Li
authors.initials: L
authors.lastname: Xiao
authors.forename: Doris M
authors.initials: DM
authors.lastname: Haverstick
authors.forename: Alison
authors.initials: A
authors.lastname: Dewald
authors.forename: Linda
authors.initials: L
authors.lastname: Columbus
authors.forename: Kimberly
authors.initials: K
authors.lastname: Kelly
authors.forename: James P
authors.initials: JP
authors.lastname: Landers
mesh_terms: D002452:Cell Count / Q000379:methods; D004578:Electron Spin Resonance Spectroscopy / Q000379:methods; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008863:Microspheres; D013601:T-Lymphocytes / Q000737:chemistry*