DNA Electroporation, Isolation and Imaging of Myofibers.
نویسندگان
چکیده
Mature muscle has a unique structure that is amenable to live cell imaging. Herein, we describe the experimental protocol for expressing fluorescently labeled proteins in the flexor digitorum brevis (FDB) muscle. Conditions have been optimized to provide a large number of high quality myofibers expressing the electroporated plasmid while minimizing muscle damage. The method employs fluorescent tags on various proteins. Combining this expression method with high resolution confocal microscopy permits live cell imaging, including imaging after laser-induced damage. Fluorescent dyes combined with imaging of fluorescently-tagged proteins provides information regarding the basic structure of muscle and its response to stimuli.
منابع مشابه
P-151: Optimal Condition of IntroducingGreen Fluorescent Protein (GFP) Geneinto The Rainbow Trout (Oncorhynchusmykiss) Sperm by Electroporation Method
Background: Many techniques have been developed to increase DNA molecules into sperms called sperm mediated gene transfer (SMGT) and one of them is electroporation. Sperm mediated gene transfer is an ideal method for mass gene transfer in organisms with external fertilization. Gene transfer is applied to rainbow trout sperm and the effects of electroporation on sperms and rate of gene uptake by...
متن کاملEvaluation of Soft Tissue Sarcoma Tumors Electrical Conductivity Anisotropy Using Diffusion Tensor Imaging for Numerical Modeling on Electroporation
Introduction: There is many ways to assessing the electrical conductivity anisotropyof a tumor. Applying the values of tissue electrical conductivity anisotropyis crucial in numerical modeling of the electric and thermal field distribution in electroporationtreatments. This study aims to calculate the tissues electrical conductivityanisotropy in patients with sarcoma tumors using diffusion tens...
متن کاملOptimal Electroporation Condition for Small Interfering RNA Transfection into MDA-MB-468 Cell Line
Background: Electroporation is a valuable tool for small interfering RNA (siRNA) delivery into cells because it efficiently transforms a wide variety of cell types. Since electroporation condition for each cell type must be determined experimentally, this study presents an optimal electroporation strategy to reproducibly and efficiently transfect MDA-MB 468 human breast cancer cell with siRNA. ...
متن کاملIn vivo time-lapse imaging of neuronal development in Xenopus.
In vivo fluorescence imaging of cells in the developing nervous system is greatly facilitated in specimens in which cells are brightly but sparsely labeled. In this article, we describe a number of techniques that can be used for delivering fluorophore to neurons in the albino Xenopus laevis tadpole. Fluorescent dye or DNA that encodes a fluorescent protein can be delivered to single cells by e...
متن کاملIsolation, Culture, and Staining of Single Myofibers.
Adult skeletal muscle regeneration is orchestrated by a specialized population of adult stem cells called satellite cells, which are localized between the basal lamina and the plasma membrane of myofibers. The process of satellite cell-activation, proliferation, and subsequent differentiation that occurs during muscle regeneration can be recapitulated ex vivo by isolation of single myofibers fr...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Journal of visualized experiments : JoVE
دوره 106 شماره
صفحات -
تاریخ انتشار 2015