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Adv Healthc Mater. 2013 Jul;2(7):965-975. doi: 10.1002/adhm.201200378. Epub 2013 Jan 20.

Surface-micromachined microfiltration membranes for efficient isolation and functional immunophenotyping of subpopulations of immune cells.

Author information

1
Department of Mechanical Engineering University of Michigan Ann Arbor, MI 48109 USA.
2
Department of Mechanical and Biomedical Engineering City University of Hong Kong, Hong Kong, China.
3
Department of Biomedical Engineering Yale University New Haven, CT 06511, USA.
4
Department of Pediatrics and Communicable Diseases University of Michigan, Ann Arbor, MI 48109, USA.
#
Contributed equally

Abstract

An accurate measurement of the immune status in patients with immune system disorders is critical in evaluating the stage of diseases and tailoring drug treatments. The functional cellular immunity test is a promising method to establish the diagnosis of immune dysfunctions. The conventional functional cellular immunity test involves measurements of the capacity of peripheral blood mononuclear cells to produce pro-inflammatory cytokines when stimulated ex vivo. However, this "bulk" assay measures the overall reactivity of a population of lymphocytes and monocytes, making it difficult to pinpoint the phenotype or real identity of the reactive immune cells involved. In this research, we develop a large surface micromachined poly-dimethylsiloxane (PDMS) microfiltration membrane (PMM) with high porosity, which is integrated in a microfluidic microfiltration platform. Using the PMM with functionalized microbeads conjugated with antibodies against specific cell surface proteins, we demonstrated rapid, efficient and high-throughput on-chip isolation, enrichment, and stimulation of subpopulations of immune cells from blood specimens. Furthermore, the PMM-integrated microfiltration platform, coupled with a no-wash homogeneous chemiluminescence assay ("AlphaLISA"), enables us to demonstrate rapid and sensitive on-chip immunophenotyping assays for subpopulations of immune cells isolated directly from minute quantities of blood samples.

KEYWORDS:

biomedical applications; cell separation; immune cell; immunophenotyping; microfluidics

PMID:
23335389
PMCID:
PMC4459734
DOI:
10.1002/adhm.201200378
[Indexed for MEDLINE]
Free PMC Article

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