Automated manipulation and sorting of single cells are challenging, when intact cells are needed for further investigations, e.g., RNA or DNA sequencing. We applied a computer controlled micropipette on a microscope admitting 80 PCR (Polymerase Chain Reaction) tubes to be filled with single cells in a cycle. Due to the Laplace pressure, fluid starts to flow out from the micropipette only above a critical pressure preventing the precise control of drop volume in the submicroliter range. We found an anomalous pressure additive to the Laplace pressure that we attribute to the evaporation of the drop. We have overcome the problem of the critical dropping pressure with sequentially operated fast fluidic valves timed with a millisecond precision. Minimum drop volume was 0.4–0.7 μl with a sorting speed of 15–20 s per cell. After picking NE-4C neuroectodermal mouse stem cells and human primary monocytes from a standard plastic Petri dish we could gently deposit single cells inside tiny drops. 94 ± 3% and 54 ± 7% of the deposited drops contained single cells for NE-4C and monocytes, respectively. 7.5 ± 4% of the drops contained multiple cells in case of monocytes. Remaining drops were empty. Number of cells deposited in a drop could be documented by imaging the Petri dish before and after sorting. We tuned the adhesion force of cells to make the manipulation successful without the application of microstructures for trapping cells on the surface. We propose that our straightforward and flexible setup opens an avenue for single cell isolation, critically needed for the rapidly growing field of single cell biology.
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25 August 2014
Research Article|
August 27 2014
Automated single cell sorting and deposition in submicroliter drops
Rita Salánki;
Rita Salánki
1Doctoral School of Molecular- and Nanotechnologies,
University of Pannonia
, Veszprém H-8200 Hungary
2Nanobiosensorics Group, Research Centre for Natural Sciences,
Institute for Technical Physics and Materials Science
, Konkoly Thege M. út 29-33. 1121 Budapest, Hungary
3Department of Biological Physics,
Eötvös University
, Pázmány Péter sétány 1A, Budapest H-1117, Hungary
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Tamás Gerecsei;
Tamás Gerecsei
3Department of Biological Physics,
Eötvös University
, Pázmány Péter sétány 1A, Budapest H-1117, Hungary
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Norbert Orgovan;
Norbert Orgovan
2Nanobiosensorics Group, Research Centre for Natural Sciences,
Institute for Technical Physics and Materials Science
, Konkoly Thege M. út 29-33. 1121 Budapest, Hungary
3Department of Biological Physics,
Eötvös University
, Pázmány Péter sétány 1A, Budapest H-1117, Hungary
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Noémi Sándor;
Noémi Sándor
4MTA-ELTE Immunology Research Group,
Eötvös University
, Budapest H-1117, Hungary
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Beatrix Péter;
Beatrix Péter
1Doctoral School of Molecular- and Nanotechnologies,
University of Pannonia
, Veszprém H-8200 Hungary
2Nanobiosensorics Group, Research Centre for Natural Sciences,
Institute for Technical Physics and Materials Science
, Konkoly Thege M. út 29-33. 1121 Budapest, Hungary
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Zsuzsa Bajtay;
Zsuzsa Bajtay
5Department of Immunology,
Eötvös University
, Budapest H-1117, Hungary
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Anna Erdei;
Anna Erdei
4MTA-ELTE Immunology Research Group,
Eötvös University
, Budapest H-1117, Hungary
5Department of Immunology,
Eötvös University
, Budapest H-1117, Hungary
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Robert Horvath;
Robert Horvath
2Nanobiosensorics Group, Research Centre for Natural Sciences,
Institute for Technical Physics and Materials Science
, Konkoly Thege M. út 29-33. 1121 Budapest, Hungary
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Bálint Szabó
Bálint Szabó
a)
2Nanobiosensorics Group, Research Centre for Natural Sciences,
Institute for Technical Physics and Materials Science
, Konkoly Thege M. út 29-33. 1121 Budapest, Hungary
3Department of Biological Physics,
Eötvös University
, Pázmány Péter sétány 1A, Budapest H-1117, Hungary
6
CellSorter Company for Innovations
, Erdőalja út 174, Budapest H-1037, Hungary
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a)
Author to whom correspondence should be addressed. Electronic mail: balintszabo1@gmail.com.
Appl. Phys. Lett. 105, 083703 (2014)
Article history
Received:
May 29 2014
Accepted:
August 10 2014
Citation
Rita Salánki, Tamás Gerecsei, Norbert Orgovan, Noémi Sándor, Beatrix Péter, Zsuzsa Bajtay, Anna Erdei, Robert Horvath, Bálint Szabó; Automated single cell sorting and deposition in submicroliter drops. Appl. Phys. Lett. 25 August 2014; 105 (8): 083703. https://doi.org/10.1063/1.4893922
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