کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5591640 1404987 2017 15 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Vitrification after multiple rounds of sample application and blotting improves particle density on cryo-electron microscopy grids
ترجمه فارسی عنوان
تصفیه پس از چندین دور از برنامه نمونه و بلوط، چگالی ذرات بر روی شبکه های میکروسکوپی یون های الکترونی
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شناسی مولکولی
چکیده انگلیسی
Single particle cryo-electron microscopy (cryoEM) is becoming widely adopted as a tool for structural characterization of biomolecules at near-atomic resolution. Vitrification of the sample to obtain a dense distribution of particles within a single field of view remains a major bottleneck for the success of such experiments. Here, we describe a simple and cost-effective method to increase the density of frozen-hydrated particles on grids with holey carbon support films. It relies on performing multiple rounds of sample application and blotting prior to plunge freezing in liquid ethane. We show that this approach is generally applicable and significantly increases particle density for a range of samples, such as small protein complexes, viruses and filamentous assemblies. The method is versatile, easy to implement, minimizes sample requirements and can enable characterization of samples that would otherwise resist structural studies using single particle cryoEM.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Structural Biology - Volume 198, Issue 1, April 2017, Pages 38-42
نویسندگان
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