Single Cell Sampling Using Micromanipulation: A Technique to Isolate Single Cells from Cancer Cell Suspension

Published: April 30, 2023

Abstract

Source: Chen S. et al. Target Cell Pre-enrichment and Whole Genome Amplification for Single Cell Downstream Characterization. J. Vis. Exp. (2018)

This video describes the technique for isolating single cells from a pool of target cells to carry out downstream analysis. In this method, a micromanipulator fitted to a microscope is used for high-precision isolation of a single target cell.

Protocol

1. Single-cell Sampling Micromanipulation ​NOTE: To allow downstream applications like whole-genome analysis (WGA), isolated single cells will be added to 2 µL of cell lysis master mix. Prepare a master mix according to the number of cells to be processed. Calculate extra volumes for loss due to pipetting and place cell lysis master mix on ice. Prepare cell lysis master mix (components of WGA kit, Table of Materials …

開示

The authors have nothing to disclose.

Materials

Phosphate Buffered Saline (1x PBS)  Thermo Fisher Scientific  10010-015
Vacutainer EDTA (or Na-heparin) tubes    BD 366450 (or 366480) Used as reaction tube for ex vivo capture of target cells
Pasteur pipettes 150 mm    Volac D810 Used for the low-volume application of cells to the C&R
Grease pen   Dako S2002 Used on glass slides to hold cell suspension in place
Axiovert M200 equipped with Mikromanipulator MMJ and CellTram vario   Zeiss/Eppendorf Use micromanipulation at hand
Microcapillaries (25 µm in diameter), CustomTip Type I    Eppendorf 930001201 Used for micromanipulating cells
0.2 mL PCR tubes  Biozym 710920

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記事を引用
Single Cell Sampling Using Micromanipulation: A Technique to Isolate Single Cells from Cancer Cell Suspension. J. Vis. Exp. (Pending Publication), e20396, doi: (2023).

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