Collection of cells for single-cell RNA sequencing using high-resolution fluorescence microscopy

Abstract

Summary: FACS sorting followed by single-cell RNA-sequencing (SORT-Seq) is a popular procedure to select cells of interest for single-cell transcriptomics. However, FACS is not suitable for measurement of subcellular distribution of fluorescence or for small samples (<1,000 cells). The VYCAP puncher system overcomes these limitations. Here, we describe a workflow to capture, image, and collect fluorescent human retina pigment epithelium cells for SORT-Seq using this system. The workflow can be used for any cell type with a diameter of ∼5–50 μm.For complete details on the use and execution of this protocol, please refer to Segeren et al. (2020).

Keywords

• Cell Biology
• Single Cell
• Genomics
• Microscopy