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Single-cell lysis for visual analysis by electron microscopy.


Kemmerling, Simon; Arnold, Stefan A; Bircher, Benjamin A; Sauter, Nora; Escobedo, Carlos; Dernick, Gregor; Hierlemann, Andreas; Stahlberg, Henning; Braun, Thomas (2013). Single-cell lysis for visual analysis by electron microscopy. Journal of Structural Biology, 183(3):467-73.

Abstract

The stochastic nature of biological systems makes the study of individual cells a necessity in systems biology. Yet, handling and disruption of single cells and the analysis of the relatively low concentrations of their protein components still challenges available techniques. Transmission electron microscopy (TEM) allows for the analysis of proteins at the single-molecule level. Here, we present a system for single-cell lysis under light microscopy observation, followed by rapid uptake of the cell lysate. Eukaryotic cells were grown on conductively coated glass slides and observed by light microscopy. A custom-designed microcapillary electrode was used to target and lyse individual cells with electrical pulses. Nanoliter volumes were subsequently aspirated into the microcapillary and dispensed onto an electron microscopy grid for TEM inspection. We show, that the cell lysis and preparation method conserves protein structures well and is suitable for visual analysis by TEM.

Abstract

The stochastic nature of biological systems makes the study of individual cells a necessity in systems biology. Yet, handling and disruption of single cells and the analysis of the relatively low concentrations of their protein components still challenges available techniques. Transmission electron microscopy (TEM) allows for the analysis of proteins at the single-molecule level. Here, we present a system for single-cell lysis under light microscopy observation, followed by rapid uptake of the cell lysate. Eukaryotic cells were grown on conductively coated glass slides and observed by light microscopy. A custom-designed microcapillary electrode was used to target and lyse individual cells with electrical pulses. Nanoliter volumes were subsequently aspirated into the microcapillary and dispensed onto an electron microscopy grid for TEM inspection. We show, that the cell lysis and preparation method conserves protein structures well and is suitable for visual analysis by TEM.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:Special Collections > SystemsX.ch
Special Collections > SystemsX.ch > Research, Technology and Development Projects > CINA
Dewey Decimal Classification:570 Life sciences; biology
Language:English
Date:2013
Deposited On:11 Sep 2013 17:53
Last Modified:05 Apr 2016 16:58
Publisher:Elsevier
ISSN:1047-8477
Publisher DOI:https://doi.org/10.1016/j.jsb.2013.06.012
PubMed ID:23816812

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