Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients | Zendy

Changfu Yao | Zendy; Stephanie A. Bora | Zendy; Peter Chen | Zendy; Helen S. Goodridge | Zendy; Sina A. Gharib | Zendy

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Sample processing and single cell RNA-sequencing of peripheral blood immune cells from COVID-19 patients

Author(s) -

Changfu Yao,

Stephanie A. Bora,

Peter Chen,

Helen S. Goodridge,

Sina A. Gharib

Publication year - 2021

Publication title -

star protocols

Language(s) - English

Resource type - Journals

ISSN - 2666-1667

DOI - 10.1016/j.xpro.2021.100582

Subject(s) - buffy coat , peripheral blood mononuclear cell , immune system , rna , virology , covid-19 , cell , cell sorting , immunology , biology , peripheral blood , computational biology , medicine , gene , flow cytometry , pathology , genetics , infectious disease (medical specialty) , in vitro , disease

Single-cell RNA sequencing (scRNA-seq) of PBMCs allows in-depth assessment of transcriptional changes in immune cells of COVID-19 patients. However, collecting, processing, and analyzing samples from COVID-19 patients pose many challenges because blood samples may contain infectious virus, identification of immune cell subtypes can be difficult, and biological interpretation of analytical results is complex. To address these issues, we describe a protocol for sample processing, sorting, methanol fixation and scRNA-seq analysis of PBMCs from frozen buffy coat samples from COVID-19 patients.

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