Isolation and counting of multiple cell types using an affinity separation device.
A simple device for the separation of cells by phenotype is described. Cells are separated/isolated using capture antibodies on a glass chip. Unlike other "sandwich" type assays, the readout is performed without labels using transmission microscopy, simplifying cell enumeration. T and B lymphocytes from cell culture or whole blood were separated using antibodies for the CD4, CD19, and CD71 antigens. The separation slides were found to reproducibly bind cells by antigen expression, allowing for accurate enumeration of mixed cell samples. Inter- and intra-device variability was evaluated, and the issue of nonspecific binding is addressed. We envision that this type of cell separation technique could be used in remote settings, as sample preparation is minimal and the analysis time is rapid (20 min from sample acquisition to final readout).
Wang
Kelong
K
Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, TX 79409-1061, USA.
Cometti
Brandon
B
Pappas
Dimitri
D
D000906:Antibodies / Q000276:immunology; D015703:Antigens, CD / Q000032:analysis; D001402:B-Lymphocytes / Q000737:chemistry; D002452:Cell Count / Q000295:instrumentation*; D002460:Cell Line; D002469:Cell Separation / Q000295:instrumentation*; D005434:Flow Cytometry / Q000295:instrumentation*; D006801:Humans; D008853:Microscopy; D013601:T-Lymphocytes / Q000737:chemistry
D000906:Antibodies; D015703:Antigens, CD