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Total Sample Conditioning and Preparation of Nanoliter Volumes for Electron Microscopy

Arnold, Stefan A. and Albiez, Stefan and Opara, Nadia and Chami, Mohamed and Schmidli, Claudio and Bieri, Andrej and Padeste, Celestino and Stahlberg, Henning and Braun, Thomas. (2016) Total Sample Conditioning and Preparation of Nanoliter Volumes for Electron Microscopy. ACS Nano, 10 (5). pp. 4981-4988.

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Official URL: http://edoc.unibas.ch/51773/

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Abstract

Electron microscopy (EM) entered a new era with the emergence of direct electron detectors and new nanocrystal electron diffraction methods. However, sample preparation techniques have not progressed and still suffer from extensive blotting steps leading to a massive loss of sample. Here, we present a simple but versatile method for the almost lossless sample conditioning and preparation of nanoliter volumes of biological samples for EM, keeping the sample under close to physiological condition. A microcapillary is used to aspirate 3-5 nL of sample. The microcapillary tip is immersed into a reservoir of negative stain or trehalose, where the sample becomes conditioned by diffusive exchange of salt and heavy metal ions or sugar molecules, respectively, before it is deposited as a small spot onto an EM grid. We demonstrate the use of the method to prepare protein particles for imaging by transmission EM and nanocrystals for analysis by electron diffraction. Furthermore, the minute sample volume required for this method enables alternative strategies for biological experiments, such as the analysis of the content of a single cell by visual proteomics, fully exploiting the single molecule detection limit of EM.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Stahlberg)
UniBasel Contributors:Stahlberg, Henning
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:American Chemical Society
ISSN:1936-0851
e-ISSN:1936-086X
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:25 Oct 2017 15:14
Deposited On:25 Oct 2017 15:14

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