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Title: Label-free quantitative lymphocyte activation profiling using microfluidic impedance cytometry
Authors: Petchakup, Chayakorn
Hutchinson, Paul Edward
Tay, Hui Min
Leong, Sheng Yuan
Li, Holden King Ho
Hou, Han Wei
Keywords: Science::Medicine
Issue Date: 2021
Source: Petchakup, C., Hutchinson, P. E., Tay, H. M., Leong, S. Y., Li, H. K. H. & Hou, H. W. (2021). Label-free quantitative lymphocyte activation profiling using microfluidic impedance cytometry. Sensors and Actuators B: Chemical, 339, 129864-.
Project: RG53/18
Journal: Sensors and Actuators B: Chemical
Abstract: Circulating lymphocytes are integral components of our adaptive immunity with emerging clinical applications in immune status monitoring in infectious diseases and cell-mediated cancer immunotherapies. Herein we present a novel impedance-based microfluidic assay for label-free lymphocyte activation profiling based on native or antigen-specific T-lymphocyte biophysical responses. Single cell impedance profiling of T-lymphocytes first revealed distinct biophysical differences in cell size and membrane electrical impedance of healthy, activated (CD3/CD28) and dead lymphocyte populations. Impedance characterization of peripheral blood mononuclear cells (PBMCs) stimulated with mitogen phytohemagglutinin (PHA) or Tuberculin Purified Protein Derivative antigen (PPD) after 24 h also showed an increase in lymphocyte cell size (∼8 to 10 μm) which corresponded to activated lymphocytes (CD69+CD137+). We next developed a spiral inertial microfluidics cell sorter integrated with coplanar electrodes for direct impedance quantification of activated lymphocytes. By removing non-activated smaller lymphocytes (< 8 μm) and employing hydrodynamic-based single stream particle focusing, we demonstrated significant enrichment of activated lymphocytes (∼11.7-fold) to electrically detect low levels of lymphocyte activation (< 5%). Finally, the developed biochip is coupled with magnetic activated cell sorting (MACS) to quantify CD4+ T-lymphocytes response in PBMCs stimulated with PPD. A differential impedance cell count ratio (stimulated/unstimulated) was defined to distinguish activated T-lymphocytes, which showed better sensitivity as compared to immunophenotyping by flow cytometry. Taken together, the integrated impedance biosensor can be further developed as a rapid multiplexed screening assay to detect antigen-specific T-lymphocyte responses to characterize host immunity and diagnosis of infectious diseases (e.g tuberculosis, dengue and COVID-19).
ISSN: 0925-4005
DOI: 10.1016/j.snb.2021.129864
Schools: School of Mechanical and Aerospace Engineering 
Lee Kong Chian School of Medicine (LKCMedicine) 
Organisations: Singapore-MIT Alliance for Research and Technology
Rights: © 2021 Elsevier B.V. All rights reserved.
Fulltext Permission: none
Fulltext Availability: No Fulltext
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