Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes
Summary: High dimensional compositional and transcriptional profiling of heterogeneous brain-infiltrating leukocytes can lead to novel biological and therapeutic discoveries. High-quality single-cell leukocyte preparations are a prerequisite for optimal single cell profiling. Here, we describe a pro...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721002446 |
| _version_ | 1819069085285089280 |
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| author | Ian H. Guldner Samantha M. Golomb Qingfei Wang Emilia Wang Siyuan Zhang |
| author_facet | Ian H. Guldner Samantha M. Golomb Qingfei Wang Emilia Wang Siyuan Zhang |
| author_sort | Ian H. Guldner |
| collection | DOAJ |
| description | Summary: High dimensional compositional and transcriptional profiling of heterogeneous brain-infiltrating leukocytes can lead to novel biological and therapeutic discoveries. High-quality single-cell leukocyte preparations are a prerequisite for optimal single cell profiling. Here, we describe a protocol for epitope and RNA-preserving dissociation of adult mouse brains and subsequent leukocyte purification and staining, which is adaptable to homeostatic and pathogenic brains. Leukocyte preparation following this protocol permits exquisite single-cell surface protein and RNA profiling in applications including CyTOF and CITE-seq.For complete details on the use and execution of this protocol, please refer to Guldner et al. (2020) and Golomb et al. (2020). |
| first_indexed | 2024-12-21T16:44:26Z |
| format | Article |
| id | doaj.art-52efb14e334549ea8bdf8a98bd490461 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-21T16:44:26Z |
| publishDate | 2021-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-52efb14e334549ea8bdf8a98bd4904612022-12-21T18:57:01ZengElsevierSTAR Protocols2666-16672021-06-0122100537Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomesIan H. Guldner0Samantha M. Golomb1Qingfei Wang2Emilia Wang3Siyuan Zhang4Department of Biological Sciences, College of Science, University of Notre Dame, Notre Dame, IN, 46556, USA; Mike and Josie Harper Cancer Research Institute, University of Notre Dame, South Bend, IN, 46617, USA; Stanford University, Stanford, CA, 94305, USA; Corresponding authorDepartment of Biological Sciences, College of Science, University of Notre Dame, Notre Dame, IN, 46556, USA; Mike and Josie Harper Cancer Research Institute, University of Notre Dame, South Bend, IN, 46617, USADepartment of Biological Sciences, College of Science, University of Notre Dame, Notre Dame, IN, 46556, USA; Mike and Josie Harper Cancer Research Institute, University of Notre Dame, South Bend, IN, 46617, USADepartment of Biological Sciences, College of Science, University of Notre Dame, Notre Dame, IN, 46556, USA; Mike and Josie Harper Cancer Research Institute, University of Notre Dame, South Bend, IN, 46617, USADepartment of Biological Sciences, College of Science, University of Notre Dame, Notre Dame, IN, 46556, USA; Mike and Josie Harper Cancer Research Institute, University of Notre Dame, South Bend, IN, 46617, USA; Indiana University Melvin and Bren Simon Cancer Center, Indianapolis, IN, 46202, USA; Corresponding authorSummary: High dimensional compositional and transcriptional profiling of heterogeneous brain-infiltrating leukocytes can lead to novel biological and therapeutic discoveries. High-quality single-cell leukocyte preparations are a prerequisite for optimal single cell profiling. Here, we describe a protocol for epitope and RNA-preserving dissociation of adult mouse brains and subsequent leukocyte purification and staining, which is adaptable to homeostatic and pathogenic brains. Leukocyte preparation following this protocol permits exquisite single-cell surface protein and RNA profiling in applications including CyTOF and CITE-seq.For complete details on the use and execution of this protocol, please refer to Guldner et al. (2020) and Golomb et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166721002446Cell isolationSingle CellFlow Cytometry/Mass CytometrySequencingRNA-seqImmunology |
| spellingShingle | Ian H. Guldner Samantha M. Golomb Qingfei Wang Emilia Wang Siyuan Zhang Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes STAR Protocols Cell isolation Single Cell Flow Cytometry/Mass Cytometry Sequencing RNA-seq Immunology |
| title | Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes |
| title_full | Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes |
| title_fullStr | Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes |
| title_full_unstemmed | Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes |
| title_short | Isolation of mouse brain-infiltrating leukocytes for single cell profiling of epitopes and transcriptomes |
| title_sort | isolation of mouse brain infiltrating leukocytes for single cell profiling of epitopes and transcriptomes |
| topic | Cell isolation Single Cell Flow Cytometry/Mass Cytometry Sequencing RNA-seq Immunology |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721002446 |
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