Cryo-focused-ion-beam applications in structural biology

2015 ◽  
Vol 581 ◽  
pp. 122-130 ◽  
Author(s):  
Alexander Rigort ◽  
Jürgen M. Plitzko
Keyword(s):  
Structural Biology ◽  
Focused Ion Beam ◽  
Ion Beam

scholarly journals Automated cryo-lamella preparation for high-throughput in-situ structural biology

2019 ◽  
Author(s):  
Genevieve Buckley ◽  
Gediminas Gervinskas ◽  
Cyntia Taveneau ◽  
Hari Venugopal ◽  
James C. Whisstock ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Structural Biology ◽  
Focused Ion Beam ◽  
Electron Tomography ◽  
Ion Beam ◽  
Automated Method ◽  
Cellular Environment ◽  
Transmission Electron

AbstractCryo-transmission electron tomography (cryo-ET) in association with cryo-focused ion beam (cryo-FIB) milling enables structural biology studies to be performed directly within the cellular environment. Cryo-preserved cells are milled and a lamella with a thickness of 200-300 nm provides an electron transparent window suitable for cryo-ET imaging. Cryo-FIB milling is an effective method, but it is a tedious and time-consuming process, which typically results in ~10 lamellae per day. Here, we introduce an automated method to reproducibly prepare cryo-lamellae on a grid and reduce the amount of human supervision. We tested the routine on cryo-preserved Saccharomyces cerevisiae and demonstrate that this method allows an increased throughput, achieving a rate of 5 lamellae/hour without the need to supervise the FIB milling. We demonstrate that the quality of the lamellae is consistent throughout the preparation and their compatibility with cryo-ET analyses.

Protocol for cryo-focused ion beam lift-out technique

2019 ◽  
Author(s):  
Miroslava Schaffer ◽  
Stefan Pfeffer ◽  
Julia Mahamid ◽  
Stephan Kleindiek ◽  
Tim Laugks ◽  
...  
Keyword(s):  
High Pressure ◽  
Structural Biology ◽  
Focused Ion Beam ◽  
Preparation Method ◽  
Electron Tomography ◽  
Ion Beam ◽  
Cell Interior ◽  
Cryo Electron Tomography ◽  
Multicellular Organisms

Abstract Cryo-focused ion beam milling of frozen hydrated cells for the production of thin lamellas in combination with cryo-electron tomography (cryo-ET) has yielded unprecedented insights into the cell interior. This method allows access to native structures deep inside cells, enabling structural studies of macromolecules in situ. However, it is only suitable for cells that can be vitrified by plunge freezing (<10 μm). Multicellular organisms and tissues are considerably thicker and high-pressure freezing is required to ensure optimal preservation. Here, we describe a preparation method for extracting lamellas from high pressure frozen samples with a new cryo-gripper tool. This in situ lift-out technique at cryo-temperatures enables cryo-ET to be performed on multicellular organisms and tissue, extending the range of applications for in situ structural biology.

Cryo-Focused Ion Beam Lamella Preparation Protocol for in Situ Structural Biology

2021 ◽  
pp. 301-322
Author(s):  
Jana Moravcová ◽  
Radka Dopitová ◽  
Matyáš Pinkas ◽  
Jiří Nováček
Keyword(s):  
Structural Biology ◽  
Focused Ion Beam ◽  
Ion Beam

Nanocomposite Polyurethane Foams Via POSS Blends

2002 ◽  
Vol 733 ◽  
Author(s):  
Brock McCabe ◽  
Steven Nutt ◽  
Brent Viers ◽  
Tim Haddad
Keyword(s):  
High Temperature ◽  
Sample Preparation ◽  
Room Temperature ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Polyurethane Foams ◽  
Development Projects ◽  
Polymer Systems ◽  
Polyurethane Resin ◽  
Military Development

AbstractPolyhedral Oligomeric Silsequioxane molecules have been incorporated into a commercial polyurethane formulation to produce nanocomposite polyurethane foam. This tiny POSS silica molecule has been used successfully to enhance the performance of polymer systems using co-polymerization and blend strategies. In our investigation, we chose a high-temperature MDI Polyurethane resin foam currently used in military development projects. For the nanofiller, or “blend”, Cp7T7(OH)3 POSS was chosen. Structural characterization was accomplished by TEM and SEM to determine POSS dispersion and cell morphology, respectively. Thermal behavior was investigated by TGA. Two methods of TEM sample preparation were employed, Focused Ion Beam and Ultramicrotomy (room temperature).

An Study of Influence of Imperfections on the Delamination of Diamond-Like Carbon Films

2002 ◽  
Vol 719 ◽  
Author(s):  
Myoung-Woon Moon ◽  
Kyang-Ryel Lee ◽  
Jin-Won Chung ◽  
Kyu Hwan Oh
Keyword(s):  
Cross Sections ◽  
Focused Ion Beam ◽  
Imaging System ◽  
Ion Beam ◽  
Carbon Films ◽  
Diamond Like Carbon ◽  
Glass Substrates ◽  
Atomic Force ◽  
Initiation And Propagation

AbstractThe role of imperfections on the initiation and propagation of interface delaminations in compressed thin films has been analyzed using experiments with diamond-like carbon (DLC) films deposited onto glass substrates. The surface topologies and interface separations have been characterized by using the Atomic Force Microscope (AFM) and the Focused Ion Beam (FIB) imaging system. The lengths and amplitudes of numerous imperfections have been measured by AFM and the interface separations characterized on cross sections made with the FIB. Chemical analysis of several sites, performed using Auger Electron Spectroscopy (AES), has revealed the origin of the imperfections. The incidence of buckles has been correlated with the imperfection length.

Low Energy Ar Ion Milling of FIB TEM Specimens from 14 nm and Future FinFET Technologies

2018 ◽  
Author(s):  
C.S. Bonifacio ◽  
P. Nowakowski ◽  
M.J. Campin ◽  
M.L. Ray ◽  
P.E. Fischione
Keyword(s):  
Field Effect Transistor ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Specimen Preparation ◽  
Ion Milling ◽  
Transmission Electron ◽  
High Resolution Tem ◽  
Effect Transistor ◽  
20 Nm ◽  
Argon Ion

Abstract Transmission electron microscopy (TEM) specimens are typically prepared using the focused ion beam (FIB) due to its site specificity, and fast and accurate thinning capabilities. However, TEM and high-resolution TEM (HRTEM) analysis may be limited due to the resulting FIB-induced artifacts. This work identifies FIB artifacts and presents the use of argon ion milling for the removal of FIB-induced damage for reproducible TEM specimen preparation of current and future fin field effect transistor (FinFET) technologies. Subsequently, high-quality and electron-transparent TEM specimens of less than 20 nm are obtained.

Automated Diagonal Slice and View Solution for 3D Device Structure Analysis

2018 ◽  
Author(s):  
Sang Hoon Lee ◽  
Jeff Blackwood ◽  
Stacey Stone ◽  
Michael Schmidt ◽  
Mark Williamson ◽  
...  
Keyword(s):  
Cross Section ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Image Data ◽  
Planar Surface ◽  
Device Structure ◽  
Cross Sectional ◽  
Device Structures ◽  
The Cross ◽  
Planar Analysis

Abstract The cross-sectional and planar analysis of current generation 3D device structures can be analyzed using a single Focused Ion Beam (FIB) mill. This is achieved using a diagonal milling technique that exposes a multilayer planar surface as well as the cross-section. this provides image data allowing for an efficient method to monitor the fabrication process and find device design errors. This process saves tremendous sample-to-data time, decreasing it from days to hours while still providing precise defect and structure data.

Novel Approach of Improving Secondary Electron Detector in FIB System

2018 ◽  
Author(s):  
Steve Wang ◽  
Jim McGinn ◽  
Peter Tvarozek ◽  
Amir Weiss
Keyword(s):  
Integrated Circuit ◽  
Secondary Electron ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Vital Role ◽  
Electron Detector ◽  
System A ◽  
Novel Approach

Abstract Secondary electron detector (SED) plays a vital role in a focused ion beam (FIB) system. A successful circuit edit requires a good effective detector. Novel approach is presented in this paper to improve the performance of such a detector, making circuit altering for the most advanced integrated circuit (IC) possible.

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