Comparison of 3D cellular imaging techniques based on scanned electron probes: Serial block face SEM vs. Axial bright-field STEM tomography

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dc.contributor.author McBride, E. L.
dc.contributor.author Rao, A.
dc.contributor.author Zhang, G.
dc.contributor.author Hoyne, J. D.
dc.contributor.author Calco, G. N.
dc.contributor.author Kuo, B. C.
dc.contributor.author He, Q.
dc.contributor.author Prince, A. A.
dc.contributor.author Pokrovskaya, I. D.
dc.contributor.author Storrie, B.
dc.contributor.author Sousa, A. A. [UNIFESP]
dc.contributor.author Aronova, M. A.
dc.contributor.author Leapman, R. D.
dc.date.accessioned 2018-07-26T12:18:38Z
dc.date.available 2018-07-26T12:18:38Z
dc.date.issued 2018
dc.identifier http://dx.doi.org/10.1016/j.jsb.2018.01.012
dc.identifier.citation Journal Of Structural Biology. San Diego, v. 202, n. 3, p. 216-228, 2018.
dc.identifier.issn 1047-8477
dc.identifier.uri http://repositorio.unifesp.br/handle/11600/45995
dc.description.abstract Microscopies based on focused electron probes allow the cell biologist to image the 3D ultrastructure of eukaryotic cells and tissues extending over large volumes, thus providing new insight into the relationship between cellular architecture and function of organelles. Here we compare two such techniques: electron tomography in conjunction with axial bright-field scanning transmission electron microscopy (BF-STEM), and serial block face scanning electron microscopy (SBF-SEM). The advantages and limitations of each technique are illustrated by their application to determining the 3D ultrastructure of human blood platelets, by considering specimen geometry, specimen preparation, beam damage and image processing methods. Many features of the complex membranes composing the platelet organelles can be determined from both approaches, although STEM tomography offers a higher similar to 3 nm isotropic pixel size, compared with similar to 5 nm for SBF-SEM in the plane of the block face and similar to 30 nm in the perpendicular direction. In this regard, we demonstrate that STEM tomography is advantageous for visualizing the platelet canalicular system, which consists of an interconnected network of narrow (similar to 50-100 nm) membranous cisternae. In contrast, SBF-SEM enables visualization of complete platelets, each of which extends similar to 2 mu m in minimum dimension, whereas BF-STEM tomography can typically only visualize approximately half of the platelet volume due to a rapid non-linear loss of signal in specimens of thickness greater than similar to 1.5 mu m. We also show that the limitations of each approach can be ameliorated by combining 3D and 2D measurements using a stereological approach. en
dc.description.sponsorship National Institute of Biomedical Imaging and Bioengineering at the National Institutes of Health in Bethesda, Maryland
dc.format.extent 216-228
dc.language.iso eng
dc.publisher Academic Press Inc Elsevier Science
dc.rights Acesso restrito
dc.subject Scanning transmission electron microscopy (STEM) en
dc.subject Electron tomography en
dc.subject Serial block face scanning electron microscopy (SBF-SEM) en
dc.subject Human blood platelets en
dc.title Comparison of 3D cellular imaging techniques based on scanned electron probes: Serial block face SEM vs. Axial bright-field STEM tomography en
dc.type Artigo
dc.description.affiliation NIBIB, Lab Cellular Imaging & Macromol Biophys, NIN, Bethesda, MD USA
dc.description.affiliation Univ Arkansas Med Sci, Dept Physiol & Biophys, Little Rock, AR 72205 USA
dc.description.affiliation Univ Fed Sao Paulo, Dept Biochem, Sao Paulo, Brazil
dc.description.affiliationUnifesp Univ Fed Sao Paulo, Dept Biochem, Sao Paulo, Brazil
dc.description.sponsorshipID NIH: R01 HL119393
dc.identifier.doi 10.1016/j.jsb.2018.01.012
dc.description.source Web of Science
dc.identifier.wos WOS:000432639000005



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