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Cells 2014, 3(4), 1102-1115;

Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays

ZellNet Consulting, Inc., 555 North Avenue, Suite 25-S, Fort Lee, NJ 07024, USA
Autoimmun Diagnostika GmbH, Ebinger Strasse 4, D-72479 Strassberg, Germany
Mabtech AB, Box 1233, SE-131 28 Nacka Strand, Sweden
Author to whom correspondence should be addressed.
Received: 8 October 2014 / Revised: 13 November 2014 / Accepted: 14 November 2014 / Published: 27 November 2014
(This article belongs to the Special Issue ELISPOT Research)
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ELISpot is one of the most commonly used immune monitoring assays, which allows the functional assessment of the immune system at the single cell level. With its outstanding sensitivity and ease of performance, the assay has recently advanced from the mere single function cell analysis to multifunctional analysis by implementing detection reagents that are labeled with fluorophores (FluoroSpot), allowing the detection of secretion patterns of two or more analytes in a single well. However, the automated evaluation of such assays presents various challenges for image analysis. Here we dissect the technical and methodological requirements for a reliable analysis of FluoroSpot assays, introduce important quality control measures and provide advice for proper interpretation of results obtained by automated imaging systems. View Full-Text
Keywords: FluoroSpot; ELISpot; immunomonitoring; image analysis; fluorochromes; fluorophores FluoroSpot; ELISpot; immunomonitoring; image analysis; fluorochromes; fluorophores

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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Janetzki, S.; Rueger, M.; Dillenbeck, T. Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays. Cells 2014, 3, 1102-1115.

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