کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8038335 1518335 2014 11 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers
ترجمه فارسی عنوان
میکروسکوپ همبستگی بالا و میکروسکوپ الکترونی با استفاده از دو نشانگر همگرایی مستقل
موضوعات مرتبط
مهندسی و علوم پایه مهندسی مواد فناوری نانو (نانو تکنولوژی)
چکیده انگلیسی
Correlative light and electron microscopy (CLEM) is an emerging technique which combines functional information provided by fluorescence microscopy (FM) with the high-resolution structural information of electron microscopy (EM). So far, correlative cryo microscopy of frozen-hydrated samples has not reached better than micrometre range accuracy. Here, a method is presented that enables the correlation between fluorescently tagged proteins and electron cryo tomography (cryoET) data with nanometre range precision. Specifically, thin areas of vitrified whole cells are examined by correlative fluorescence cryo microscopy (cryoFM) and cryoET. Novel aspects of the presented cryoCLEM workflow not only include the implementation of two independent electron dense fluorescent markers to improve the precision of the alignment, but also the ability of obtaining an estimate of the correlation accuracy for each individual object of interest. The correlative workflow from plunge-freezing to cryoET is detailed step-by-step for the example of locating fluorescence-labelled adenovirus particles trafficking inside a cell.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Ultramicroscopy - Volume 143, August 2014, Pages 41-51
نویسندگان
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