On account of its excellent resolution and high throughput, cryoSEM imaging has recently seen resurgence. In this work, we report on the development of cryogenic triple ion gun milling (CryoTIGM™), a broad ion beam milling technique for cryo-planing of vitrified, "frozen-hydrated" specimens. We find that sections prepared with CryoTIGM™ are smooth over exceptionally large areas (~700,000 µm2), and reveal ultrastructural details in similar or better quality than freeze-fractured samples.
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http://dx.doi.org/10.1017/S143192761501510X | DOI Listing |
BMC Biol
June 2023
Institute of Physics, School of Basic Sciences, École Polytechnique Fédérale de Lausanne, Lausanne, CH-1015, Switzerland.
Background: The development of nanoscale secondary ion mass spectrometry (NanoSIMS) has revolutionized the study of biological tissues by enabling, e.g., the visualization and quantification of metabolic processes at subcellular length scales.
View Article and Find Full Text PDFMicrosc Microanal
December 2015
Department of Materials Science and Engineering, Northwestern University,2220 Campus Drive,Evanston,IL 60208,USA.
On account of its excellent resolution and high throughput, cryoSEM imaging has recently seen resurgence. In this work, we report on the development of cryogenic triple ion gun milling (CryoTIGM™), a broad ion beam milling technique for cryo-planing of vitrified, "frozen-hydrated" specimens. We find that sections prepared with CryoTIGM™ are smooth over exceptionally large areas (~700,000 µm2), and reveal ultrastructural details in similar or better quality than freeze-fractured samples.
View Article and Find Full Text PDFJ Struct Biol
November 2015
Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried 82152, Germany. Electronic address:
Cryo-electron tomography provides 3D views of cellular architecture with molecular resolution. A principal limitation of cryo-transmission electron microscopy performed on cells or tissues is the accessible specimen thickness. Recently it has been shown that cryo-focused ion beam milling of plunge-frozen eukaryotic cells can produce homogeneously thin, distortion free lamellas for cryo-electron tomography.
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