title: On-chip immune cell activation and subsequent time-resolved magnetic bead-based cytokine detection.
abstract: Cytokine profiling and immunophenotyping offer great potential for understanding many disease mechanisms, personalized diagnosis, and immunotherapy. Here, we demonstrate a time-resolved detection of cytokine from a single cell cluster using an in situ magnetic immune assay. An array of triple-layered microfluidic chambers was fabricated to enable simultaneous cell culture under perfusion flow and detection of the induced cytokines at multiple time-points. Each culture chamber comprises three fluidic compartments which are dedicated to, cell culture, perfusion and immunoassay. The three compartments are separated by porous membranes, which allow the diffusion of fresh nutrient from the perfusion compartment into the cell culture compartment and cytokines secretion from the cell culture compartment into the immune assay compartment. This structure hence enables capturing the released cytokines without disturbing the cell culture and without minimizing benefit gain from perfusion. Functionalized magnetic beads were used as a solid phase carrier for cytokine capturing and quantification. The cytokines released from differential stimuli were quantified in situ in non-differentiated U937 monocytes and differentiated macrophages.
authors.affiliation: Institute of Microelectronics, Agency for Science Technology and Research (A*STAR), 2 Fusionopolis Way, #08-02 Innovis Tower, Singapore, 138634, Singapore. kongsupholp@ime.a-star.edu.sg.
authors.forename: Patthara
authors.initials: P
authors.lastname: Kongsuphol
authors.affiliation: Institute of Microelectronics, Agency for Science Technology and Research (A*STAR), 2 Fusionopolis Way, #08-02 Innovis Tower, Singapore, 138634, Singapore.
authors.forename: Yunxiao
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Institute of Microelectronics, Agency for Science Technology and Research (A*STAR), 2 Fusionopolis Way, #08-02 Innovis Tower, Singapore, 138634, Singapore.
authors.forename: Qasem
authors.initials: Q
authors.lastname: Ramadan
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D016207:Cytokines / Q000032:analysis*; D006801:Humans; D007118:Immunoassay / Q000295:instrumentation*; D056656:Lab-On-A-Chip Devices; D059346:Magnets; D008863:Microspheres; D013997:Time Factors
chemical_list: D016207:Cytokines
keywords: Cytokines; Immune cell activation; Immune cells; Magnetic beads assay; Microfluidic; Time-resolved assay