Articles by Anastasia Syntychaki in JoVE
Miniaturized Sample Preparation for Transmission Electron Microscopy Claudio Schmidli*1,2, Luca Rima*1, Stefan A. Arnold*1,2, Thomas Stohler1, Anastasia Syntychaki1, Andrej Bieri1, Stefan Albiez1, Kenneth N. Goldie1, Mohamed Chami3, Henning Stahlberg1, Thomas Braun1 1Center for Cellular Imaging and NanoAnalytics (C-CINA), Biozentrum, University of Basel, 2Swiss Nanoscience Institute, University of Basel, 3BioEM lab, Biozentrum, University of Basel An instrument and methods for the preparation of nanoliter-sized sample volumes for transmission electron microscopy is presented. No paper-blotting steps are required, thus avoiding the detrimental consequences this can have for proteins, significantly reducing sample loss and enabling the analysis of single cell lysate for visual proteomics.
Other articles by Anastasia Syntychaki on PubMed
Blotting-free and Lossless Cryo-electron Microscopy Grid Preparation from Nanoliter-sized Protein Samples and Single-cell Extracts Journal of Structural Biology. 03, 2017 | Pubmed ID: 27864160 We present a sample preparation method for cryo-electron microscopy (cryo-EM) that requires only 3-20nL of sample to prepare a cryo-EM grid, depending on the protocol used. The sample is applied and spread on the grid by a microcapillary. The procedure does not involve any blotting steps, and real-time monitoring allows the water film thickness to be assessed and decreased to an optimum value prior to vitrification. We demonstrate that the method is suitable for high-resolution cryo-EM and will enable alternative electron microscopy approaches, such as single-cell visual proteomics.
Miniaturizing EM Sample Preparation: Opportunities, Challenges, and "Visual Proteomics" Proteomics. Mar, 2018 | Pubmed ID: 29441686 This review compares and discusses conventional versus miniaturized specimen preparation methods for transmission electron microscopy (TEM). The progress brought by direct electron detector cameras, software developments and automation have transformed transmission cryo-electron microscopy (cryo-EM) and made it an invaluable high-resolution structural analysis tool. In contrast, EM specimen preparation has seen very little progress in the last decades and is now one of the main bottlenecks in cryo-EM. Here, we discuss the challenges faced by specimen preparation for single particle EM, highlight current developments, and show the opportunities resulting from the advanced miniaturized and microfluidic sample grid preparation methods described, such as visual proteomics and time-resolved cryo-EM studies.