Flow Imaging of the Inflammasome: Evaluating ASC Speck Characteristics and Caspase-1 Activity.

Autor:	Nagar A; Program in Innate Immunity, Division of Infectious Diseases and Immunology, School of Medicine, University of Massachusetts, Worcester, MA, USA., Harton JA; Department of Immunology and Microbial Disease, Albany Medical College, Albany, NY, USA. hartonj@amc.edu.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2023; Vol. 2635, pp. 185-202.
DOI:	10.1007/978-1-0716-3020-4_11
Abstrakt:	Examining inflammasome-associated speck structures is one of the most preferred and easiest ways to evaluate inflammasome activation. Microscopy-based evaluation of specks is preferable, but this approach is time-consuming and limited to small sample sizes. Speck-containing cells can also be quantitated by a flow cytometric method, time of flight inflammasome evaluation (TOFIE). However, TOFIE cannot perform single-cell analysis such as simultaneously visualizing ASC specks and caspase-1 activity, their location, and physical characteristics. Here we describe the application of an imaging flow cytometry-based approach that overcomes these limitations. Inflammasome and Caspase-1 Activity Characterization and Evaluation (ICCE) is a high-throughput, single-cell, rapid image analysis utilizing the Amnis ImageStream X instrument with over 99.5% accuracy. ICCE quantitatively and qualitatively characterizes the frequency, area, and cellular distribution of ASC specks and caspase-1 activity in mouse and human cells. (© 2023. Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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