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Automated large volume sample preparation for vEM.
Stempinski, Erin S; Pagano, Lucas; Riesterer, Jessica L; Adamou, Steven K; Thibault, Guillaume; Song, Xubo; Chang, Young Hwan; López, Claudia S.
Afiliação
  • Stempinski ES; Multiscale Microscopy Core, Oregon Health & Science University, Portland, OR, United States.
  • Pagano L; Knight Cancer Institute-CEDAR, Oregan Health & Science University, Portland, OR, United States.
  • Riesterer JL; Multiscale Microscopy Core, Oregon Health & Science University, Portland, OR, United States; Knight Cancer Institute-CEDAR, Oregan Health & Science University, Portland, OR, United States.
  • Adamou SK; Multiscale Microscopy Core, Oregon Health & Science University, Portland, OR, United States.
  • Thibault G; Knight Cancer Institute-CEDAR, Oregan Health & Science University, Portland, OR, United States; Department of Biomedical Engineering, Oregon Health & Science University, Portland, OR, United States.
  • Song X; Knight Cancer Institute-CEDAR, Oregan Health & Science University, Portland, OR, United States.
  • Chang YH; Department of Biomedical Engineering, Oregon Health & Science University, Portland, OR, United States.
  • López CS; Multiscale Microscopy Core, Oregon Health & Science University, Portland, OR, United States; Department of Biomedical Engineering, Oregon Health & Science University, Portland, OR, United States; Pacific Northwest Center for Cryo-EM, Oregon Health & Science University, Portland, OR, Unit
Methods Cell Biol ; 177: 1-32, 2023.
Article em En | MEDLINE | ID: mdl-37451763
ABSTRACT
New developments in electron microscopy technology, improved efficiency of detectors, and artificial intelligence applications for data analysis over the past decade have increased the use of volume electron microscopy (vEM) in the life sciences field. Moreover, sample preparation methods are continuously being modified by investigators to improve final sample quality, increase electron density, combine imaging technologies, and minimize the introduction of artifacts into specimens under study. There are a variety of conventional bench protocols that a researcher can utilize, though most of these protocols require several days. In this work, we describe the utilization of an automated specimen processor, the mPrep™ ASP-2000™, to prepare samples for vEM that are compatible with focused ion beam scanning electron microscopy (FIB-SEM), serial block face scanning electron microscopy (SBF-SEM), and array tomography (AT). The protocols described here aimed for methods that are completed in a much shorter period of time while minimizing the exposure of the operator to hazardous and toxic chemicals and improving the reproducibility of the specimens' heavy metal staining, all without compromising the quality of the data acquired using backscattered electrons during SEM imaging. As a control, we have included a widely used sample bench protocol and have utilized it as a comparator for image quality analysis, both qualitatively and using image quality analysis metrics.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Inteligência Artificial / Imageamento Tridimensional Idioma: En Revista: Methods Cell Biol Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Inteligência Artificial / Imageamento Tridimensional Idioma: En Revista: Methods Cell Biol Ano de publicação: 2023 Tipo de documento: Article País de afiliação: Estados Unidos