Conjugate Multimode Heat Transfer Analysis of Cryogenic EXLO Manipulation.

In this study, a conjugate radiation/conduction multimode heat transfer analysis of cryogenic focused ion beam (FIB) milling steps necessary for producing ex situ lift out specimens under cryogenic conditions (cryo-EXLO) is performed. Using finite volume for transient heat conduction and enclosure theory for radiation heat transfer, the analysis shows that as long as the specimen is attached or touching the FIB side wall trenches, the specimen will remain vitreous indefinitely, while actively cooled at liquid nitrogen (LN2) temperatures. To simulate the time needed to perform a transfer step to move the bulk sample containing the FIB-thinned specimen from the cryo-FIB to the cryo-EXLO cryostat, the LN2 temperature active cooling is turned off after steady-state conditions are reached and the specimen is monitored over time until the critical devitrification temperature is reached. Under these conditions, the sample will remain vitreous for >3 min, which is more than enough time needed to perform the cryo-transfer step from the FIB to the cryostat, which takes only ∼10 s. Cryo-transmission electron microscopy images of a manipulated cryo-EXLO yeast specimen prepared with cryo-FIB corroborates the heat transfer analysis.

Mechanism of FIB-Induced Phase Transformation in Austenitic Steel.

The transformation of unstable austenite to ferrite or α' martensite as a result of exposure to Xe+ or Ga+ ions at room temperature was studied in a 304 stainless steel casting alloy. Controlled Xe+ and Ga+ ion beam exposures of the 304 were carried out at a variety of beam/sample geometries. It was found that both Ga+ and Xe+ ion irradiation resulted in the transformation of the austenite to either ferrite or α' martensite. In this paper, we will refer to the transformation product as a BCC phase. The crystallographic orientation of the transformed area was controlled by the orientation of the austenite grain and was consistent with either the Nishiyama­Wasserman or the Kurdjumov­Sachs orientation relationships. On the basis of the Xe+ and Ga+ ion beam exposures, the transformation is not controlled by the chemical stabilization of the BCC phase by the ion species, but is a result of the disorder caused by the ion-induced recoil motion and subsequent return of the disordered region to a more energetically favorable phase.

Theory and New Applications of Ex Situ Lift Out.

The ex situ lift out (EXLO) adhesion forces are reviewed and new applications of EXLO for focused ion beam (FIB)-prepared specimens are described. EXLO is used to manipulate electron transparent specimens on microelectromechanical systems carrier devices designed for in situ electron microscope analysis. A new patented grid design without a support film is described for EXLO. This new slotted grid design provides a surface for holding the specimen in place and also allows for post lift out processing. Specimens may be easily manipulated into a backside orientation to reduce FIB curtaining artifacts with this slotted grid. Large EXLO specimens can be manipulated from Xe+ plasma FIB prepared specimens. Finally, applications of EXLO and manipulation of FIB specimens using a vacuum probe lift out method are shown. The vacuum probe provides more control for placing specimens on the new slotted grids and also allows for easy manipulation into a backside configuration.

Replacing a failed mini-implant with a miniplate to prevent interruption during orthodontic treatment.

INTRODUCTION: When mini-implants fail during orthodontic treatment, there is a need to have a backup plan to either replace the failed implant in the adjacent interradicular area or wait for the bone to heal before replacing the mini-implant. We propose a novel way to overcome this problem by replacement with a miniplate so as not to interrupt treatment or prolong treatment time. METHODS: The indications, advantages, efficacy, and procedures for switching from a mini-implant to a miniplate are discussed. Two patients who required replacement of failed mini-implants are presented. In the first patient, because of the proximity of the buccal vestibule to the mini-implant, it was decided to replace the failed mini-implant by an I-shaped C-tube miniplate. In the second patient, radiolucencies were found around the failed mini-implants, making the adjacent alveolar bone unavailable for immediate placement of another mini-implant. In addition, the maxillary sinus pneumatization was expanded deeply into the interradicular spaces; this further mandated an alternative placement site. One failed mini-implant was examined under a scanning electron microscope for bone attachment. RESULTS: Treatment was completed in both patients after replacement with miniplates without interrupting the treatment mechanics or prolonging the treatments. Examination under the scanning electron microscope showed partial bone growth into the coating pores and titanium substrate interface even after thorough cleaning and sterilization. CONCLUSIONS: Replacement with a miniplate is a viable solution for failed mini-implants during orthodontic treatment. The results from microscopic evaluation of the failed mini-implant suggest that stringent guidelines are needed for recycling used mini-implants.

Two-dimensional and 3-dimensional analysis of bone/dental implant interfaces with the use of focused ion beam and electron microscopy.

PURPOSE: To assess the usefulness of a dual-beam focused ion beam (FIB) and scanning electron microscope (SEM) instrument and FIB-based transmission electron microscopy (TEM) specimen preparation techniques to characterize bone/dental implant interfaces. MATERIALS AND METHODS: The FIB was used to site specifically polished cross-sections for direct FIB, SEM, and TEM imaging of bone osseointegration into a Nobel Biocare TiUnite failed dental implant (Nobel Biocare, Yorba Linda, CA). RESULTS: Bone was observed to grow into the porous structure of the coating, yielding direct evidence of a mechanical locking mechanism of the bone/implant interface. Multiple SEM images obtained from sequential FIB cross-sections were reconstructed into 3-dimensional tomograms that showed partial and full bone growth into the porous structure of the TiUnite coating. Sections thinned by FIB techniques were observed by transmission electron microscope (TEM) and related methods. Conventional bright field TEM showed that the coating, which was more than 2 microm thick, consisted of a nanocrystalline and porous structure. High-resolution TEM (HRTEM) showed the presence within the bone of hydroxyapatite crystallites that measured approximately 7 nm. TEM images showed that the bone does not form an intimate and homogenous interface with the implant coating in all regions. X-ray energy dispersive spectrometer (XEDS) line scans that used scanning TEM (STEM) methods showed interdiffusion of Ti, P, and Ca between the bone and the coating where intimate bone/coating contact was observed, suggesting that chemical bonding also exists within this interface. CONCLUSIONS: FIB methods for SEM and TEM were used to characterize bone/implant surfaces.

Site-specific 3D imaging of cells and tissues with a dual beam microscope.

Current approaches to 3D imaging at subcellular resolution using confocal microscopy and electron tomography, while powerful, are limited to relatively thin and transparent specimens. Here we report on the use of a new generation of dual beam electron microscopes capable of site-specific imaging of the interior of cellular and tissue specimens at spatial resolutions about an order of magnitude better than those currently achieved with optical microscopy. The principle of imaging is based on using a focused ion beam to create a cut at a designated site in the specimen, followed by viewing the newly generated surface with a scanning electron beam. Iteration of these two steps several times thus results in the generation of a series of surface maps of the specimen at regularly spaced intervals, which can be converted into a three-dimensional map of the specimen. We have explored the potential of this sequential "slice-and-view" strategy for site-specific 3D imaging of frozen yeast cells and tumor tissue, and establish that this approach can identify the locations of intracellular features such as the 100 nm-wide yeast nuclear pore complex. We also show that 200 nm thick sections can be generated in situ by "milling" of resin-embedded specimens using the ion beam, providing a valuable alternative to manual sectioning of cells and tissues using an ultramicrotome. Our results demonstrate that dual beam imaging is a powerful new tool for cellular and subcellular imaging in 3D for both basic biomedical and clinical applications.

Site-specific Transmission Electron Microscope Characterization of Micrometer-sized Particles Using the Focused Ion Beam Lift-out Technique.

Micrometer sized particles have been studied to show that a high-quality transmission electron microscope (TEM) specimen can be produced, without the use of embedding media, from a site-specific region of chosen particles using the focused ion beam (FIB) lift-out (LO) technique. The uniqueness of this technique is that site-specific TEM LO specimens may be obtained from particles and from regions which are smaller than the conventional approximately 10-20 &mgr;m x 5 &mgr;m x approximately 0.1 &mgr;m dimensions of the LO specimen. The innovative FIB LO procedures are described in detail and TEM images of electron transparent specimens obtained from specific micrometer-sized particles are presented.