دورية أكاديمية
أعرض في EDS

Automated transmission-mode scanning electron microscopy (tSEM) for large volume analysis at nanoscale resolution.

التفاصيل البيبلوغرافية
العنوان: Automated transmission-mode scanning electron microscopy (tSEM) for large volume analysis at nanoscale resolution.
المؤلفون: Kuwajima M; Center for Learning and Memory, The University of Texas at Austin, Austin, Texas, United States of America., Mendenhall JM, Lindsey LF, Harris KM
المصدر: PloS one [PLoS One] 2013; Vol. 8 (3), pp. e59573. Date of Electronic Publication: 2013 Mar 26.
نوع المنشور: Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Public Library of Science Country of Publication: United States NLM ID: 101285081 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1932-6203 (Electronic) Linking ISSN: 19326203 NLM ISO Abbreviation: PLoS One Subsets: MEDLINE
أسماء مطبوعة: Original Publication: San Francisco, CA : Public Library of Science
مواضيع طبية MeSH: Microscopy, Electron, Scanning/*methods , Nanotechnology/*methods, Animals ; Artifacts ; Automation ; Brain/ultrastructure ; Cost-Benefit Analysis ; Elasticity ; Image Processing, Computer-Assisted ; Lenses ; Microscopy, Electron, Scanning/economics ; Microscopy, Electron, Scanning/instrumentation ; Nanotechnology/economics ; Nanotechnology/instrumentation ; Rats
مستخلص: Transmission-mode scanning electron microscopy (tSEM) on a field emission SEM platform was developed for efficient and cost-effective imaging of circuit-scale volumes from brain at nanoscale resolution. Image area was maximized while optimizing the resolution and dynamic range necessary for discriminating key subcellular structures, such as small axonal, dendritic and glial processes, synapses, smooth endoplasmic reticulum, vesicles, microtubules, polyribosomes, and endosomes which are critical for neuronal function. Individual image fields from the tSEM system were up to 4,295 µm(2) (65.54 µm per side) at 2 nm pixel size, contrasting with image fields from a modern transmission electron microscope (TEM) system, which were only 66.59 µm(2) (8.160 µm per side) at the same pixel size. The tSEM produced outstanding images and had reduced distortion and drift relative to TEM. Automated stage and scan control in tSEM easily provided unattended serial section imaging and montaging. Lens and scan properties on both TEM and SEM platforms revealed no significant nonlinear distortions within a central field of ∼100 µm(2) and produced near-perfect image registration across serial sections using the computational elastic alignment tool in Fiji/TrakEM2 software, and reliable geometric measurements from RECONSTRUCT™ or Fiji/TrakEM2 software. Axial resolution limits the analysis of small structures contained within a section (∼45 nm). Since this new tSEM is non-destructive, objects within a section can be explored at finer axial resolution in TEM tomography with current methods. Future development of tSEM tomography promises thinner axial resolution producing nearly isotropic voxels and should provide within-section analyses of structures without changing platforms. Brain was the test system given our interest in synaptic connectivity and plasticity; however, the new tSEM system is readily applicable to other biological systems.
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معلومات مُعتمدة: NS074644 United States NS NINDS NIH HHS; R01 NS074644 United States NS NINDS NIH HHS; R56 MH095980 United States MH NIMH NIH HHS; R01 MH104319 United States MH NIMH NIH HHS; MH095980 United States MH NIMH NIH HHS; R01 NS021184 United States NS NINDS NIH HHS; NS021184 United States NS NINDS NIH HHS; R01 MH095980 United States MH NIMH NIH HHS; R37 NS021184 United States NS NINDS NIH HHS
تواريخ الأحداث: Date Created: 20130405 Date Completed: 20130930 Latest Revision: 20220317
رمز التحديث: 20240628
مُعرف محوري في PubMed: PMC3608656
DOI: 10.1371/journal.pone.0059573
PMID: 23555711
قاعدة البيانات: MEDLINE
الوصف
تدمد:1932-6203
DOI:10.1371/journal.pone.0059573