scholarly journals Tb3+-doped LaF3nanocrystals for correlative cathodoluminescence electron microscopy imaging with nanometric resolution in focused ion beam-sectioned biological samples

Nanoscale ◽  
2017 ◽  
Vol 9 (13) ◽  
pp. 4383-4387 ◽  
Author(s):  
K. Keevend ◽  
M. Stiefel ◽  
A. L. Neuer ◽  
M. T. Matter ◽  
A. Neels ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Biological Samples ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Microscopy Imaging

scholarly journals Protocol for preparation of heterogeneous biological samples for 3D electron microscopy: a case study for insects

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexey A. Polilov ◽  
Anastasia A. Makarova ◽  
Song Pang ◽  
C. Shan Xu ◽  
Harald Hess
Keyword(s):  
Electron Microscopy ◽  
Biological Samples ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Three Dimensional ◽  
Entire Volume ◽  
Simple Protocol ◽  
3D Electron Microscopy ◽  
3D Em

AbstractModern morphological and structural studies are coming to a new level by incorporating the latest methods of three-dimensional electron microscopy (3D-EM). One of the key problems for the wide usage of these methods is posed by difficulties with sample preparation, since the methods work poorly with heterogeneous (consisting of tissues different in structure and in chemical composition) samples and require expensive equipment and usually much time. We have developed a simple protocol allows preparing heterogeneous biological samples suitable for 3D-EM in a laboratory that has a standard supply of equipment and reagents for electron microscopy. This protocol, combined with focused ion-beam scanning electron microscopy, makes it possible to study 3D ultrastructure of complex biological samples, e.g., whole insect heads, over their entire volume at the cellular and subcellular levels. The protocol provides new opportunities for many areas of study, including connectomics.

scholarly journals Revealing the cell-material interface with nanometer resolution by FIB-SEM

2017 ◽  
Author(s):  
Francesca Santoro ◽  
Wenting Zhao ◽  
Lydia-Marie Joubert ◽  
Liting Duan ◽  
Jan Schnitker ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Titanium Implants ◽  
Chemical Compositions ◽  
Material Interface ◽  
Microscopy Imaging ◽  
Substrate Removal ◽  
Cell Material

The interface between biological cells and non-biological surfaces profoundly influences cellular activities, chronic tissue responses, and ultimately the success of medical implants. Materials in contact with cells can be plastics, metal, ceramics or other synthetic materials, and their surfaces vary widely in chemical compositions, stiffness, topography and levels of roughness. To understand the molecular mechanism of how cells and tissues respond to different materials, it is of critical importance to directly visualize the cell-material interface at the relevant length scale of nanometers. Conventional ultrastructural analysis by transmission electron microscopy (TEM) often requires substrate removal before microtome sectioning, which is not only challenging for most substrates but also can cause structural distortions of the interface. Here, we present a new method for in situ examination of the cell-to-material interface at any desired cellular location, based on focused-ion beam milling and scanning electron microscopy imaging (FIB-SEM). This method involves a thin-layer plastification procedure that preserves adherent cells as well as enhances the contrast of biological specimen. We demonstrate that this unique procedure allows the visualization of cell-to-material interface and intracellular structures with 10nm resolution, compatible with a variety of materials and surface topographies, and capable of volume and multi-directional imaging. We expect that this method will be very useful for studies of cell-to-material interactions and also suitable for in vivo studies such as examining osteoblast adhesion and new bone formation in response to titanium implants.

Applications of Ion Microscopy and In Situ Electron Microscopy to the Study of Electronic Materials and Devices

1998 ◽  
Vol 4 (3) ◽  
pp. 308-316 ◽  
Author(s):  
R. Hull ◽  
J. Demarest ◽  
D. Dunn ◽  
E.A. Stach ◽  
Q. Yuan
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Focused Ion Beam ◽  
Electronic Materials ◽  
Ion Beam ◽  
Microscopy Imaging ◽  
Transmission Electron ◽  
In Situ Electron Microscopy ◽  
Ion Microscopy

We discuss the application of ion microscopy and in situ electron microscopy to the study of electronic and optical materials and devices. We demonstrate how the combination of in situ transmission electron microscopy and focused ion beam microscopy provides new avenues for the study for such structures, enabling extension of these techniques to the study of dopant distributions, nanoscale stresses, three-dimensional structural and chemical reconstruction, and real-time evolution of defect microstructure. We also discuss in situ applications of thermal, mechanical, electrical, and optical stresses during transmission electron microscopy imaging.

scholarly journals From Light Microscopy to Analytical Scanning Electron Microscopy (SEM) and Focused Ion Beam (FIB)/SEM in Biology: Fixed Coordinates, Flat Embedding, Absolute References

2018 ◽  
Vol 24 (5) ◽  
pp. 526-544 ◽  
Author(s):  
Manja Luckner ◽  
Gerhard Wanner
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Biological Samples ◽  
Focused Ion Beam ◽  
Single Cells ◽  
Ion Beam ◽  
Target Cells ◽  
Wide Range ◽  
Flat Embedding ◽  
Scanning Electron

AbstractCorrelative light and electron microscopy (CLEM) has been in use for several years, however it has remained a costly method with difficult sample preparation. Here, we report a series of technical improvements developed for precise and cost-effective correlative light and scanning electron microscopy (SEM) and focused ion beam (FIB)/SEM microscopy of single cells, as well as large tissue sections. Customized coordinate systems for both slides and coverslips were established for thin and ultra-thin embedding of a wide range of biological specimens. Immobilization of biological samples was examined with a variety of adhesives. For histological sections, a filter system for flat embedding was developed. We validated ultra-thin embedding on laser marked slides for efficient, high-resolution CLEM. Target cells can be re-located within minutes in SEM without protracted searching and correlative investigations were reduced to a minimum of preparation steps, while still reaching highest resolution. The FIB/SEM milling procedure is facilitated and significantly accelerated as: (i) milling a ramp becomes needless, (ii) significant re-deposition of milled material does not occur; and (iii) charging effects are markedly reduced. By optimizing all technical parameters FIB/SEM stacks with 2 nm iso-voxels were achieved over thousands of sections, in a wide range of biological samples.

Three-Dimensional Semiconductor Device Investigation Using Focused Ion Beam and Scanning Electron Microscopy Imaging (FIB/SEM Tomography)

2013 ◽  
Vol 19 (1) ◽  
pp. 85-92 ◽  
Author(s):  
K. Lepinay ◽  
F. Lorut
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Focused Ion Beam ◽  
Semiconductor Device ◽  
Ion Beam ◽  
Three Dimensional ◽  
Turnaround Time ◽  
Volume Reconstruction ◽  
Microscopy Imaging ◽  
Scanning Electron

AbstractThree-dimensional focused ion beam/scanning electron microscopy (FIB/SEM tomography) is currently an important technique to characterize in 3D a complex semiconductor device or a specific failure. However, the industrial context demands low turnaround time making the technique less useful. To make it more attractive, the following study focuses on a specific methodology going from sample preparation to the final volume reconstruction to reduce the global time analysis while keeping reliable results. The FIB/SEM parameters available will be first analyzed to acquire a relevant dataset in a reasonable time (few hours). Then, a new alignment strategy based on specific alignment marks [using tetraethoxylisane (TEOS) and Pt deposition] is proposed to improve the volume reconstruction speed. These points combined represent a considerable improvement regarding the reliability of the results and the time consumption (gain of factor 3). This method is then applied to various case studies illustrating the benefits of the FIB/SEM tomography technique such as the precise identification of the origin of 3D defects, or the capability to perform a virtual top-down deprocessing on soft material not possible by any mechanical solution.

scholarly journals Oxygen plasma focused ion beam scanning electron microscopy for biological samples

2018 ◽  
Author(s):  
Gorelick Sergey ◽  
Korneev Denis ◽  
Handley Ava ◽  
Gervinskas Gediminas ◽  
Oorschot Viola ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Sample Preparation ◽  
Biological Samples ◽  
Oxygen Plasma ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Light And Electron Microscopy ◽  
Beam Scanning ◽  
Scanning Electron

AbstractOver the past decade, gallium Focused Ion Beam-Scanning Electron Microscopy (FIB-SEM) has been established as a key technology for cellular tomography. The utility of this approach, however, is severely limited both by throughput and the limited selection of compatible sample preparation protocols. Here, we address these limitations and present oxygen plasma FIB (O-PFIB) as a new and versatile tool for cellular FIB-SEM tomography. Oxygen displays superior resin compatibility to other ion beams and produces curtain-free surfaces with minimal polishing. Our novel approach permits more flexible sample preparation and 30% faster data collection when compared to using gallium ion sources. We demonstrate this alternative FIB is applicable to a variety of embedding procedures and biological samples including brain tissue and whole organisms. Finally, we demonstrate the use of O-PFIB to produce targeted FIB-SEM tomograms through fiducial free en-block correlative light and electron microscopy.

TEM Sample Preparation Tricks for Advanced DRAMs

2015 ◽  
Author(s):  
Ching Shan Sung ◽  
Hsiu Ting Lee ◽  
Jian Shing Luo
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Sample Preparation ◽  
Integrated Circuit ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Tem Analysis ◽  
Cross Sectional ◽  
Transmission Electron ◽  
Characterization Of Materials

Abstract Transmission electron microscopy (TEM) plays an important role in the structural analysis and characterization of materials for process evaluation and failure analysis in the integrated circuit (IC) industry as device shrinkage continues. It is well known that a high quality TEM sample is one of the keys which enables to facilitate successful TEM analysis. This paper demonstrates a few examples to show the tricks on positioning, protection deposition, sample dicing, and focused ion beam milling of the TEM sample preparation for advanced DRAMs. The micro-structures of the devices and samples architectures were observed by using cross sectional transmission electron microscopy, scanning electron microscopy, and optical microscopy. Following these tricks can help readers to prepare TEM samples with higher quality and efficiency.

Fin Level Defect Isolation Inside a FinFET Using Electron Beam Alteration of Current Flow

2017 ◽  
Author(s):  
H.J. Ryu ◽  
A.B. Shah ◽  
Y. Wang ◽  
W.-H. Chuang ◽  
T. Tong
Keyword(s):  
Electron Microscopy ◽  
Electron Beam ◽  
Focused Ion Beam ◽  
Current Flow ◽  
Ion Beam ◽  
The Body ◽  
Complete Understanding ◽  
Root Cause ◽  
Transmission Electron ◽  
Defect Isolation

Abstract When failure analysis is performed on a circuit composed of FinFETs, the degree of defect isolation, in some cases, requires isolation to the fin level inside the problematic FinFET for complete understanding of root cause. This work shows successful application of electron beam alteration of current flow combined with nanoprobing for precise isolation of a defect down to fin level. To understand the mechanism of the leakage, transmission electron microscopy (TEM) slice was made along the leaky drain contact (perpendicular to fin direction) by focused ion beam thinning and lift-out. TEM image shows contact and fin. Stacking fault was found in the body of the silicon fin highlighted by the technique described in this paper.

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