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Clin Immunol. 2017 Apr;177:70-75. doi: 10.1016/j.clim.2016.03.004. Epub 2016 Mar 3.

Quantification of natural killer cell polarization and visualization of synaptic granule externalization by imaging flow cytometry.

Author information

1
Rice University, Houston, TX 77005, USA.
2
Center for Human Immunobiology, Texas Children's Hospital and Baylor College of Medicine, Houston, TX 77030, USA.
#
Contributed equally

Abstract

Defining immunological mechanisms underlying NK cell biology is crucial for the treatment and prevention of immune deficiency and malignancy. The limited availability of human biological specimens presents a challenge to the study of human immunobiology. The use of high throughput, multi-parametric assays will not only aid in the definition and diagnosis of complex human immune disorders affecting NK cell function but also advance NK cell biology through population-based assessment of molecular signaling. In an effort to garner the most information from limited numbers of human cells, we designed a quantitative method to study NK cell function using imaging flow cytometry (IFC), which combines multiparametric flow cytometry and fluorescence microscopy. Specifically, we developed IFC as a tool to measure polarization and secretion of lytic granules at the immunological synapse formed between an NK cell and a susceptible target. We have further validated our approach through quantitative comparison with high-resolution confocal microscopy. We show that IFC can be used as a quantitative, high throughput measure of NK cell biological function possessing greater dimensionality than standard flow cytometry.

KEYWORDS:

Degranulation; Flow cytometry; Imaging flow cytometry; Immunological synapse; Microscopy; Natural killer cell

PMID:
26948929
PMCID:
PMC5010793
DOI:
10.1016/j.clim.2016.03.004
[Indexed for MEDLINE]
Free PMC Article

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