Work

Highly multiplexed imaging of single cells using a high-throughput cyclic immunofluorescence method

Year: 2015

Type: article

Abstract: Single-cell analysis reveals aspects of cellular physiology not evident from population-based studies, particularly in the case of highly multiplexed methods such as mass cytometry (CyTOF) able to correlate the levels of multiple signalling, differentiation and cell fate markers. Immunofluorescence (IF) microscopy adds information on cell morphology and the microenvironment that are not obtained using flow-based techniques, but the multiplicity of conventional IF is limited. This has motivated development of imaging methods that require specialized instrumentation, exotic reagents or proprietary protocols that are difficult to reproduce in most laboratories. Here we report a public-domain method for achieving high multiplicity single-cell IF using cyclic immunofluorescence (CycIF), a simple and versatile procedure in which four-colour staining alternates with chemical inactivation of fluorophores to progressively build a multichannel image. Because CycIF uses standard reagents and instrumentation and is no more expensive than conventional IF, it is suitable for high-throughput assays and screening applications.

Source: Nature Communications

DOI: 10.1038/ncomms9390

Authors: Jia-Ren Lin, Mohammad Fallahi-Sichani, Peter K. Sorger

Institution: Harvard University

Language: English

Topic: Cell Image Analysis Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

Open Access status: gold

Estimated APC paid: $6,990

Funders: Life Sciences Research Foundation, National Institutes of Health, National Heart, Lung, and Blood Institute, National Institute of General Medical Sciences