Quantitation of IRF3 nuclear translocation in heterogeneous cellular populations from cervical tissue using imaging flow cytometry

Radiana T. Trifonova, Natalie Barteneva

Research output: Chapter in Book/Report/Conference proceedingChapter

2 Citations (Scopus)

Abstract

Imaging flow cytometry (IFC) has become a powerful tool for studying the activation of transcriptional factors in heterogeneous cell populations in high-content imaging mode. With considerable interest to the clinical development of IFC, the question becomes how we can accelerate its application to solid tissues. We developed the first IFC-based procedure to quantify the nuclear translocation of interferon regulatory factor (IRF) 3, an important measure of induction of type I interferon antiviral response, in primary human immune cells including in solid tissues. After tissue digestion and protocol optimization by spectral flow cytometry, cell suspension is stained for intracellular IRF3 and acquired by IFC. Image analysis is performed using an optimized nuclear mask and similarity score parameter to correlate the location of IRF3 staining and a nuclear dye. The technique measures IRF3 activation at a single cell level and can detect small changes in the percent of activated cells providing objective quantitative data for statistical analysis.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages125-153
Number of pages29
DOIs
Publication statusPublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1745
ISSN (Print)1064-3745

Fingerprint

Flow Cytometry
Population
Interferon Regulatory Factor-3
Interferon Type I
Statistical Data Interpretation
Masks
Transcriptional Activation
Antiviral Agents
Digestion
Suspensions
Coloring Agents
Staining and Labeling

Keywords

  • Cellular heterogeneity
  • Imaging flow cytometry
  • IRF3
  • Nucleocytoplasmic translocation
  • Signal transduction
  • Solid tissue

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Trifonova, R. T., & Barteneva, N. (2018). Quantitation of IRF3 nuclear translocation in heterogeneous cellular populations from cervical tissue using imaging flow cytometry. In Methods in Molecular Biology (pp. 125-153). (Methods in Molecular Biology; Vol. 1745). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7680-5_8

Quantitation of IRF3 nuclear translocation in heterogeneous cellular populations from cervical tissue using imaging flow cytometry. / Trifonova, Radiana T.; Barteneva, Natalie.

Methods in Molecular Biology. Humana Press Inc., 2018. p. 125-153 (Methods in Molecular Biology; Vol. 1745).

Research output: Chapter in Book/Report/Conference proceedingChapter

Trifonova, RT & Barteneva, N 2018, Quantitation of IRF3 nuclear translocation in heterogeneous cellular populations from cervical tissue using imaging flow cytometry. in Methods in Molecular Biology. Methods in Molecular Biology, vol. 1745, Humana Press Inc., pp. 125-153. https://doi.org/10.1007/978-1-4939-7680-5_8
Trifonova, Radiana T. ; Barteneva, Natalie. / Quantitation of IRF3 nuclear translocation in heterogeneous cellular populations from cervical tissue using imaging flow cytometry. Methods in Molecular Biology. Humana Press Inc., 2018. pp. 125-153 (Methods in Molecular Biology).
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