Academic Journal
Analysis of cell surface and intranuclear markers on non-stimulated human PBMC using mass cytometry
| العنوان: | Analysis of cell surface and intranuclear markers on non-stimulated human PBMC using mass cytometry |
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| المؤلفون: | Dzangué-Tchoupou, Gaëlle, Corneau, Aurélien, Blanc, Catherine, Benveniste, Olivier, Allenbach, Yves |
| المساهمون: | Centre de recherche en Myologie – U974 SU-INSERM, Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Plateforme Cytométrie Pitié-Salpêtrière (LUMIC-CYPS), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Unité Mixte de Service d'Imagerie et de Cytométrie CHU Saint-Antoine (UMS LUMIC), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), CHU Pitié-Salpêtrière AP-HP, Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU) |
| المصدر: | ISSN: 1932-6203. |
| بيانات النشر: | HAL CCSD Public Library of Science |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | [SDV]Life Sciences [q-bio] |
| الوصف: | International audience ; Mass cytometry is a powerful tool that allows simultaneous analysis of more than 37 markers at the single cell level. Mass cytometry is of particular interest in the identification of a wide variety of cell phenotypes in autoimmune diseases. Moreover, cells can be labelled with palladium isotopes and pooled before staining (barcoding). Nevertheless, immunolo-gists often face an important problem concerning the choice of markers to be included in a panel. This problem arises due to the incompatibility of different buffers used for the fixation and permeabilization of cells with various cell surface epitopes. In this study, we used a panel of 27 markers (19 surface markers and 8 intranuclear markers) to demonstrate disparities in the detection of cell surface antigens when comparing different buffers to stain unsti-mulated peripheral blood mononuclear cells. These disparities range from mild differences to very important differences in population frequencies depending on the buffers. Finally, we demonstrate the harmful effects of permeabilization prior to barcoding on the detection of some cell surface antigens. Here, we optimize a protocol that is suitable to use when targeting a large panel including both cell surface and intranuclear markers on unstimulated human peripheral blood mononuclear cells. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| Relation: | hal-01774882; https://hal.sorbonne-universite.fr/hal-01774882; https://hal.sorbonne-universite.fr/hal-01774882/document; https://hal.sorbonne-universite.fr/hal-01774882/file/journal.pone.0194593.pdf |
| DOI: | 10.1371/journal.pone.0194593 |
| الاتاحة: | https://hal.sorbonne-universite.fr/hal-01774882 https://hal.sorbonne-universite.fr/hal-01774882/document https://hal.sorbonne-universite.fr/hal-01774882/file/journal.pone.0194593.pdf https://doi.org/10.1371/journal.pone.0194593 |
| Rights: | http://creativecommons.org/licenses/by/ ; info:eu-repo/semantics/OpenAccess |
| رقم الانضمام: | edsbas.65B802D |
| قاعدة البيانات: | BASE |
| DOI: | 10.1371/journal.pone.0194593 |
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