Rapid Microfluidic Isolation Of Virally Infected Primary Bronchial Epithelial Cells For Single-Cell Rna Sequencing

Single-cell RNA sequencing (scRNA-seq) of the bronchial epithelium enables examination of cellular subtypes and their responses to viral infections. Here, an optimized method for the isolation of virally infected primary bronchial epithelial cells using a commercially available microfluidic device is presented. Using this method single cells can be rapidly isolated with minimal equipment available in most laboratories. Isolation can be carried out inside biological safety cabinets, permitting the use of virally infected cells. Both cell-line and primary cells isolated using the device retained sufficient RNA integrity for the generation of short-read sequencing-compatible cDNA libraries to facilitate scRNA-seq.METHOD SUMMARY A method for the isolation of virally infected single primary bronchial epithelial cells using a microfluidic device is presented. Using this protocol single cells can be rapidly isolated with minimal laboratory equipment and within biological safety cabinets for downstream scRNA-seq.