کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8340231 1541221 2016 12 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Nanometer resolved single-molecule colocalization of nuclear factors by two-color super resolution microscopy imaging
ترجمه فارسی عنوان
نانومتر با استفاده از دو رنگ تصویربرداری با میکروسکوپ با وضوح بالا حلقوی تک رنگ مولکولی عوامل هسته ای را حل کرد
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
چکیده انگلیسی
In order to study the detailed assembly and regulation mechanisms of complex structures and machineries in the cell, simultaneous in situ observation of all the individual interacting components should be achieved. Multi-color Single-Molecule Localization Microscopy (SMLM) is ideally suited for these quantifications. Here, we build on previous developments and thoroughly discuss a protocol for two-color SMLM combining PALM and STORM, including sample preparation details, image acquisition and data postprocessing analysis. We implement and evaluate a recently proposed colocalization analysis method (aCBC) that allows single-molecule colocalization quantification with the potential of revealing fine, nanometer-scaled, structural details of multicomponent complexes. Finally, using a doubly-labeled nuclear factor (Beaf-32) in Drosophila S2 cells we experimentally validate the colocalization quantification algorithm, highlight its advantages and discuss how using high molecular weight fluorescently labeled tags compromises colocalization precision in two-color SMLM experiments.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Methods - Volume 105, 1 August 2016, Pages 44-55
نویسندگان
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