Assembly and Imaging Set up of PIE-Scope
| Autor: | Sergey Gorelick, David Dierickx, Genevieve Buckley, James Whisstock, Alex Marco |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: | |
| Zdroj: | Bio-Protocol, Vol 10, Iss 19 (2020) |
| Druh dokumentu: | article |
| ISSN: | 2331-8325 12263559 |
| DOI: | 10.21769/BioProtoc.3768 |
| Popis: | Cryo-Electron Tomography (cryo-ET) is a method that enables resolving the structure of macromolecular complexes directly in the cellular environment. However, sample preparation for in situ Cryo-ET is labour-intensive and can require both cryo-lamella preparation through cryo-Focused Ion Beam (FIB) milling and correlative light microscopy to ensure that the event of interest is present in the lamella. Here, we present an integrated cryo-FIB and light microscope setup called the Photon Ion Electron microscope (PIE-scope) that enables direct and rapid isolation of cellular regions containing protein complexes of interest. The PIE-scope can be retrofitted on existing microscopes, although the drawings we provide are meant to work on ThermoFisher DualBeams with small mechanical modifications those can be adapted on other brands. |
| Databáze: | Directory of Open Access Journals |
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