Focused ion beam/scanning electron microscopy characterization of cell behavior on polymer micro-/nanopatterned substrates: A study of cell–substrate interactions

Martínez, Elena; Engel, Elisabeth; López‐Iglesias, Carmen; Mills, Christopher A.; Planell, Josep A.; Samitier, J.

doi:10.1016/j.micron.2006.12.003

Cited by 60 publications

(48 citation statements)

References 24 publications

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“…Exploring such continuous layer-by-layer sectioning in real time regime, it is possible to obtain information about internal three-dimensional structure of different polymer objects, as it is routinely applied for sectioning of biopolymer cellular organelles. [6] Modern SEM devices are widely applied in studies of polymer nanostructures, e.g., of microphase separation domains in thin and ultrathin block copolymer films, due to high spatial resolution together with the possibility to eliminate sputtering by conductive material and thus to preserve original surface structure to be visualised. In ref.…”

Section: Comparison With Scanning Electron Microscopy (Sem)mentioning

confidence: 99%

Scanning Force Microscopy as Applied to Conformational Studies in Macromolecular Research

Gallyamov

2011

Macromol. Rapid Commun.

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The modern state of SFM research on polymer nano-objects including single chains is discussed in comparison with other similar high-resolution microscopy techniques. The range of problems to be solved preferentially by SFM is highlighted. Promising methodology to describe quantitatively the morphology of macromolecular objects is proposed. The main benefits of this algorithm seem to be the apparent mathematical correctness as well as the possibility to estimate errors and the confidence of the numbers obtained. Special attention is paid to the dynamic observations of conformational transitions on a substrate in real time regime. This approach allows one to realise direct control of the adsorbed macromolecules by means of exposure to different vapours. Driving forces of the vapour-induced reorganisation are discussed.

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Section: Comparison With Scanning Electron Microscopy (Sem)mentioning

confidence: 99%

Scanning Force Microscopy as Applied to Conformational Studies in Macromolecular Research

Gallyamov

2011

Macromol. Rapid Commun.

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“…Our study is comparable with findings that have been described previously when FIB/SEM techniques were used to analyze such cell-to-surface interfaces. [20,32] Both groups used slightly different sample preparation techniques: critical point-dried and freeze-dried, respectively, but obtained very similar image quality and detail structures. In particular, when cellto-substrate interfaces of cell monolayers cultured on glass or metal surface were analyzed, surface damage due to uncontrollable Gallium-ion implementation, amorphization, material deposition, melting of material ( [29] and references therein) could be neglected whereas the so called ''curtain effect'' was observable (here: especially in Fig.…”

Section: B185mentioning

confidence: 99%

“…Very few studies address this topic using critical point-dried or freeze-dried cells on flat surfaces. [20,32] These studies demonstrated specifically selected sub-cellular structures for detailed characterization at the submicron scale by in situ high precision milling. [20,32] Here, we compare visualization of the cell-to material interface by conventional TEM sample preparation of neuronal networks cultured on a flat metal surface with resin-embedded in situ cross sections performed by FIB-milling.…”

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confidence: 95%

“…[20,32] These studies demonstrated specifically selected sub-cellular structures for detailed characterization at the submicron scale by in situ high precision milling. [20,32] Here, we compare visualization of the cell-to material interface by conventional TEM sample preparation of neuronal networks cultured on a flat metal surface with resin-embedded in situ cross sections performed by FIB-milling. The study also demonstrates the possibility of selecting cell-types and cellular sub-structures in critical point-dried neuronal networks on flat metal surfaces as well as the usefulness of serial sectioning at preselected regions of interest.…”

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confidence: 95%

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Imaging of Cell‐to‐Material Interfaces by SEM after in situ Focused Ion Beam Milling on Flat Surfaces and Complex 3D‐Fibrous Structures

2009

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“…These studies include elemental analysis [13] and preparation of samples for TEM. Study of interfaces between implants and other biological material (cells, tissue, bone) make up another major category [e.g 14,15,16,17], and the effect of nanoparticles on cells is also being studied [18]. Examples of the wide variety of biological specimens studied with the aid of FIB include yeast [4], chromosomes [19], atherosclerotic tissue [20], brain [7], and other organs [21].…”

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confidence: 99%

Focused Ion Beam Applications in Biology

Marko

2010

Microsc Microanal

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This tutorial is intended for biologists interested in how focused ion beam (FIB) instruments might be used in their research. It will consist of a basic introduction to FIB technology, a description of the most relevant techniques used in biology, and a selection of application examples.A FIB instrument is similar to an SEM, but produces a beam of positive ions (usually gallium) instead of electrons [1]. The ions, much heaver than electrons, can be focused by electrostatic lenses into a spot on the specimen that is a few nanometers in diameter. At beam currents of tens of picoamps or a few nanoamps, specimen material can be sputtered away to a depth of about 10 to 100 nm with each pass of the beam. This allows precision cutting and shaping of the specimen. Many modern FIB instruments include an SEM on the same vacuum chamber, the two-column "FIB-SEM". The use of SEM makes it more practical to image the specimen after cutting, without further damage.The FIB is used to prepare cross-sections or to open "windows" in the specimen for an internal view by SEM, and for making specimens for TEM. When done properly, heterogeneous hard/soft materials can be milled with a smooth surface. Frozen biological material can also be FIB-milled and observed by SEM [2] or transferred to the TEM [3].Use of FIB for TEM preparation takes three forms: (1) Thinning a portion of the specimen to TEM transparency (the "H-bar" method); this can be done at room temperature or under cryo conditions; (2) preparing a cylindrical specimen that is particularly suited to electron tomography [e.g. 4]; and (3) extracting a thin "lamella" from within a specimen and placing it on a TEM specimen holder (the "lift-out" method") [5].Probably the FIB technique of most general impact in biology is "FIB tomography", which does not involve TEM. Here, specimens are prepared by embedding in resin, after osmium fixation and subsequent en-bloc heavy-metal staining [6]. After mechanical trimming to the area of interest, the surface is alternately FIB-milled and imaged by SEM. At each milling step, about 5-20 nm of material is removed, and then SEM an image is recorded using backscattered electron imaging. The x-y resolution can be 5 nm or greater. This iterative process can proceed under software control, and it can run for days, yielding a relatively highresolution 3-D reconstruction over a large area [7,8,9]. All of the above techniques have been used extensively for over a decade in materials science, and almost as long, although with far less frequency, in biology. The first application of FIB in biology was in 1993, in which FIB was used to cut into a mite in order to image internal features [10]. Dental research represents the single largest number of publications so far [e.g.

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Focused ion beam/scanning electron microscopy characterization of cell behavior on polymer micro-/nanopatterned substrates: A study of cell–substrate interactions

Cited by 60 publications

References 24 publications

Scanning Force Microscopy as Applied to Conformational Studies in Macromolecular Research

Scanning Force Microscopy as Applied to Conformational Studies in Macromolecular Research

Imaging of Cell‐to‐Material Interfaces by SEM after in situ Focused Ion Beam Milling on Flat Surfaces and Complex 3D‐Fibrous Structures

Focused Ion Beam Applications in Biology

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