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

ABSTRACT: 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).

SUBMITTER: Segeren HA

PROVIDER: S-EPMC8353340 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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

Segeren Hendrika A HA Andree Kiki C KC Oomens Lisa L Westendorp Bart B

STAR protocols 20210805 3

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 a ...[more]

PMID: 34401784

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