Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays
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 multifunct...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2014-11-01
|
| Series: | Cells |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2073-4409/3/4/1102 |
| _version_ | 1797708771684777984 |
|---|---|
| author | Sylvia Janetzki Markus Rueger Tomas Dillenbeck |
| author_facet | Sylvia Janetzki Markus Rueger Tomas Dillenbeck |
| author_sort | Sylvia Janetzki |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-12T06:27:28Z |
| format | Article |
| id | doaj.art-d7d18c11188a43a29d618714b364f74f |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-12T06:27:28Z |
| publishDate | 2014-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-d7d18c11188a43a29d618714b364f74f2023-09-03T01:53:00ZengMDPI AGCells2073-44092014-11-01341102111510.3390/cells3041102cells3041102Stepping up ELISpot: Multi-Level Analysis in FluoroSpot AssaysSylvia Janetzki0Markus Rueger1Tomas Dillenbeck2ZellNet Consulting, Inc., 555 North Avenue, Suite 25-S, Fort Lee, NJ 07024, USAAutoimmun Diagnostika GmbH, Ebinger Strasse 4, D-72479 Strassberg, GermanyMabtech AB, Box 1233, SE-131 28 Nacka Strand, SwedenELISpot 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.http://www.mdpi.com/2073-4409/3/4/1102FluoroSpotELISpotimmunomonitoringimage analysisfluorochromesfluorophores |
| spellingShingle | Sylvia Janetzki Markus Rueger Tomas Dillenbeck Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays Cells FluoroSpot ELISpot immunomonitoring image analysis fluorochromes fluorophores |
| title | Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays |
| title_full | Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays |
| title_fullStr | Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays |
| title_full_unstemmed | Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays |
| title_short | Stepping up ELISpot: Multi-Level Analysis in FluoroSpot Assays |
| title_sort | stepping up elispot multi level analysis in fluorospot assays |
| topic | FluoroSpot ELISpot immunomonitoring image analysis fluorochromes fluorophores |
| url | http://www.mdpi.com/2073-4409/3/4/1102 |
| work_keys_str_mv | AT sylviajanetzki steppingupelispotmultilevelanalysisinfluorospotassays AT markusrueger steppingupelispotmultilevelanalysisinfluorospotassays AT tomasdillenbeck steppingupelispotmultilevelanalysisinfluorospotassays |