Development of a Miniature Cytometry Platform for Capture and Characterization of Leukocytes

Our laboratory is interested in using protein (antibody) microarrays for capture and immunophenotyping of leukocytes (T-lymphocytes) from whole blood.  In contrast to traditional flow cytometry-based approaches, our cytometry platform employs arrays of T-lymphocyte- specific antibodies (Abs) to capture CD4 and CD8 T-cells onto specific Ab spots for “solid-phase” immunophenotyping.  Microfluidic conduit is employed to enclose antibody microarrays in order to minimize the sample volume and to expose cells to a controlled shear stress.  This miniature cytometry platform has been employed to successfully capture pure (>95%) CD4+ and CD8+ T-lymphocyte subsets from whole blood.  Furthermore, the proportions of CD4+ and CD8+ residing on their respective Ab spots closely correlate to flow cytometry-based analysis of T-cell subets in a blood sample. We are currently increasing functionality of this cytometry platform by incorporating cytokine assays into the protein microarrays for investigating cytokine profiles of the captured T-cells.  This device is envisioned as a novel immunology tool with applications in HIV monitoring and diagnosis where characterization of numbers and function of T-cells is critically important.
(Researchers involved on this project: He Zhu, Gulnaz Stybayeva, David Dyer)

T-lymphocytes from human blood isolated on antibody microarrays. (A) SEM micrograph of T-cells on antibody spots. (B) Immunofluorescently stained antibody spots with captured T-cells.  (C) Immunofluorescent staining of T-cells captured on an antibody spot shows that cells are CD4+ T-cells.