Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis
In the CellSearch® system, blood is immunomagnetically enriched for epithelial cell adhesion molecule (EpCAM) expression and cells are stained with the nucleic acid dye 4′6-diamidino-2-phenylindole (DAPI), Cytokeratin-PE (CK), and CD45-APC. Only DAPI+/CK+ objects are presented to t...
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| Format: | Article |
| Language: | English |
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MDPI AG
2018-10-01
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| Series: | Cancers |
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| Online Access: | http://www.mdpi.com/2072-6694/10/10/377 |
| _version_ | 1797715901075685376 |
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| author | Sanne de Wit Leonie L. Zeune T. Jeroen N. Hiltermann Harry J. M. Groen Guus van Dalum Leon W. M. M. Terstappen |
| author_facet | Sanne de Wit Leonie L. Zeune T. Jeroen N. Hiltermann Harry J. M. Groen Guus van Dalum Leon W. M. M. Terstappen |
| author_sort | Sanne de Wit |
| collection | DOAJ |
| description | In the CellSearch® system, blood is immunomagnetically enriched for epithelial cell adhesion molecule (EpCAM) expression and cells are stained with the nucleic acid dye 4′6-diamidino-2-phenylindole (DAPI), Cytokeratin-PE (CK), and CD45-APC. Only DAPI+/CK+ objects are presented to the operator to identify circulating tumor cells (CTC) and the identity of all other cells and potential undetected CTC remains unrevealed. Here, we used the open source imaging program Automatic CTC Classification, Enumeration and PhenoTyping (ACCEPT) to analyze all DAPI+ nuclei in EpCAM-enriched blood samples obtained from 192 metastatic non-small cell lung cancer (NSCLC) patients and 162 controls. Significantly larger numbers of nuclei were detected in 300 patient samples with an average and standard deviation of 73,570 ± 74,948, as compared to 359 control samples with an average and standard deviation of 4191 ± 4463 (p < 0.001). In patients, only 18% ± 21% and in controls 23% ± 15% of the nuclei were identified as leukocytes or CTC. Adding CD16-PerCP for granulocyte staining, the use of an LED as the light source for CD45-APC excitation and plasma membrane staining obtained with wheat germ agglutinin significantly improved the classification of EpCAM-enriched cells, resulting in the identification of 94% ± 5% of the cells. However, especially in patients, the origin of the unidentified cells remains unknown. Further studies are needed to determine if undetected EpCAM+/DAPI+/CK-/CD45- CTC is present among these cells. |
| first_indexed | 2024-03-12T08:13:40Z |
| format | Article |
| id | doaj.art-35baedf17f27451db2263ca49e5c43fd |
| institution | Directory Open Access Journal |
| issn | 2072-6694 |
| language | English |
| last_indexed | 2024-03-12T08:13:40Z |
| publishDate | 2018-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cancers |
| spelling | doaj.art-35baedf17f27451db2263ca49e5c43fd2023-09-02T19:00:11ZengMDPI AGCancers2072-66942018-10-01101037710.3390/cancers10100377cancers10100377Classification of Cells in CTC-Enriched Samples by Advanced Image AnalysisSanne de Wit0Leonie L. Zeune1T. Jeroen N. Hiltermann2Harry J. M. Groen3Guus van Dalum4Leon W. M. M. Terstappen5Department of Medical Cell BioPhysics, University of Twente, 7522 NH Enschede, The NetherlandsDepartment of Medical Cell BioPhysics, University of Twente, 7522 NH Enschede, The NetherlandsDepartment of Pulmonary Diseases, University Medical Center Groningen, University of Groningen, 9713 GZ Groningen, The NetherlandsDepartment of Pulmonary Diseases, University Medical Center Groningen, University of Groningen, 9713 GZ Groningen, The NetherlandsDepartment of General, Visceral and Pediatric Surgery, University Hospital of the Heinrich-Heine-University Düsseldorf, 40225 Düsseldorf, GermanyDepartment of Medical Cell BioPhysics, University of Twente, 7522 NH Enschede, The NetherlandsIn the CellSearch® system, blood is immunomagnetically enriched for epithelial cell adhesion molecule (EpCAM) expression and cells are stained with the nucleic acid dye 4′6-diamidino-2-phenylindole (DAPI), Cytokeratin-PE (CK), and CD45-APC. Only DAPI+/CK+ objects are presented to the operator to identify circulating tumor cells (CTC) and the identity of all other cells and potential undetected CTC remains unrevealed. Here, we used the open source imaging program Automatic CTC Classification, Enumeration and PhenoTyping (ACCEPT) to analyze all DAPI+ nuclei in EpCAM-enriched blood samples obtained from 192 metastatic non-small cell lung cancer (NSCLC) patients and 162 controls. Significantly larger numbers of nuclei were detected in 300 patient samples with an average and standard deviation of 73,570 ± 74,948, as compared to 359 control samples with an average and standard deviation of 4191 ± 4463 (p < 0.001). In patients, only 18% ± 21% and in controls 23% ± 15% of the nuclei were identified as leukocytes or CTC. Adding CD16-PerCP for granulocyte staining, the use of an LED as the light source for CD45-APC excitation and plasma membrane staining obtained with wheat germ agglutinin significantly improved the classification of EpCAM-enriched cells, resulting in the identification of 94% ± 5% of the cells. However, especially in patients, the origin of the unidentified cells remains unknown. Further studies are needed to determine if undetected EpCAM+/DAPI+/CK-/CD45- CTC is present among these cells.http://www.mdpi.com/2072-6694/10/10/377circulating tumor cellsCellSearch®EpCAMleukocytesACCEPTdeep Learningclassificationimage analysisnon-small cell lung cancer |
| spellingShingle | Sanne de Wit Leonie L. Zeune T. Jeroen N. Hiltermann Harry J. M. Groen Guus van Dalum Leon W. M. M. Terstappen Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis Cancers circulating tumor cells CellSearch® EpCAM leukocytes ACCEPT deep Learning classification image analysis non-small cell lung cancer |
| title | Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis |
| title_full | Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis |
| title_fullStr | Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis |
| title_full_unstemmed | Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis |
| title_short | Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis |
| title_sort | classification of cells in ctc enriched samples by advanced image analysis |
| topic | circulating tumor cells CellSearch® EpCAM leukocytes ACCEPT deep Learning classification image analysis non-small cell lung cancer |
| url | http://www.mdpi.com/2072-6694/10/10/377 |
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