Optimized protocol for mouse footpad immune cell isolation for single-cell RNA sequencing and flow cytometry.
STAR Protoc
; 4(3): 102409, 2023 Sep 15.
Article
em En
| MEDLINE
| ID: mdl-37402171
ABSTRACT
Single-cell RNA sequencing (scRNA-seq) requires the preparation of a highly viable single-cell suspension to get reliable sequencing results. Here, we present a protocol for isolating mouse footpad leukocytes while maintaining high viability. We describe steps for footpad collection, enzymatic tissue dissociation, leukocyte isolation and purification, and cell fixation and preservation. We then detail combinatorial barcoding, library preparation, scRNA-seq, and data analysis. Cells can be used to generate a complete molecular atlas at the single cell level.
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MEDLINE
Assunto principal:
Leucócitos
Limite:
Animals
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article