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 …
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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 |
