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

Hendrika A. Segeren; Kiki C. Andree; Lisa Oomens; Bart Westendorp

doi:10.1016/j.xpro.2021.100718

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

Hendrika A. Segeren^*, Kiki C. Andree, Lisa Oomens, Bart Westendorp

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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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).

Original language	English
Article number	100718
Journal	STAR Protocols
Volume	2
Issue number	3
Early online date	5 Aug 2021
DOIs	https://doi.org/10.1016/j.xpro.2021.100718
Publication status	Published - 17 Sept 2021
Externally published	Yes

Keywords

Cell Biology
Genomics
Microscopy
Single Cell

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10.1016/j.xpro.2021.100718Licence: CC BY

ArticleFinal published version, 2.79 MBLicence: CC BY

Cite this

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title = "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).",

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doi = "10.1016/j.xpro.2021.100718",

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AU - Segeren, Hendrika A.

AU - Andree, Kiki C.

AU - Oomens, Lisa

AU - Westendorp, Bart

N1 - Funding Information: This work is financially supported by KWF Kankerbestrijding (Dutch Cancer Society, project grant 11941-2018-II ) and ZonMW (grant 91116011 ). Further financial support was provided by research infrastructure grants from Utrecht Life Sciences to the Single Cell Analysis Center and the Center for Cell Imaging. Publisher Copyright: © 2021 The Author(s)

PY - 2021/9/17

Y1 - 2021/9/17

N2 - 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).

AB - 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).

KW - Cell Biology

KW - Genomics

KW - Microscopy

KW - Single Cell

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

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Keywords

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