کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8038274 | 1518332 | 2014 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Whole-cell imaging of the budding yeast Saccharomyces cerevisiae by high-voltage scanning transmission electron tomography
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کلمات کلیدی
ADFHAADFMPLorganelleHVEM - WHOTem - این استElectron tomography - توموگرافی الکترونیStem - ساقهYeast - مخمرhigh-angle annular dark-field - میدان دایره ای بزرگ زاویه دارscanning transmission electron microscopy - میکروسکوپ الکترونی انتقال اسکنHigh-voltage electron microscopy - میکروسکوپ الکترونی با ولتاژ بالاHigh voltage electron microscopy - میکروسکوپ الکترونی بالاTransmission electron microscopy - میکروسکوپ الکترونی عبوری
موضوعات مرتبط
مهندسی و علوم پایه
مهندسی مواد
فناوری نانو (نانو تکنولوژی)
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: Whole-cell imaging of the budding yeast Saccharomyces cerevisiae by high-voltage scanning transmission electron tomography Whole-cell imaging of the budding yeast Saccharomyces cerevisiae by high-voltage scanning transmission electron tomography](/preview/png/8038274.png)
چکیده انگلیسی
Electron tomography using a high-voltage electron microscope (HVEM) provides three-dimensional information about cellular components in sections thicker than 1 μm, although in bright-field mode image degradation caused by multiple inelastic scattering of transmitted electrons limit the attainable resolution. Scanning transmission electron microscopy (STEM) is believed to give enhanced contrast and resolution compared to conventional transmission electron microscopy (CTEM). Samples up to 1 μm in thickness have been analyzed with an intermediate-voltage electron microscope because inelastic scattering is not a critical limitation, and probe broadening can be minimized. Here, we employed STEM at 1 MeV high-voltage to extend the useful specimen thickness for electron tomography, which we demonstrate by a seamless tomographic reconstruction of a whole, budding Saccharomyces cerevisiae yeast cell, which is ~3 μm in thickness. High-voltage STEM tomography, especially in the bright-field mode, demonstrated sufficiently enhanced contrast and intensity, compared to CTEM tomography, to permit segmentation of major organelles in the whole cell. STEM imaging also reduced specimen shrinkage during tilt-series acquisition. The fidelity of structural preservation was limited by cytoplasmic extraction, and the spatial resolution was limited by the relatively large convergence angle of the scanning probe. However, the new technique has potential to solve longstanding problems of image blurring in biological specimens beyond 1 μm in thickness, and may facilitate new research in cellular structural biology.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Ultramicroscopy - Volume 146, November 2014, Pages 39-45
Journal: Ultramicroscopy - Volume 146, November 2014, Pages 39-45
نویسندگان
Kazuyoshi Murata, Masatoshi Esaki, Teru Ogura, Shigeo Arai, Yuta Yamamoto, Nobuo Tanaka,