Focused ion beam/scanning electron microscopy studies of Porcellio scaber (Isopoda, Crustacea) digestive gland epithelium cells

Drobne, Damjana; Milani, M.; Zrimec, Alexis; Berden-Zrimec, Maja; Tatti, Francesco; Drašlar, Kazimir

doi:10.1002/sca.4950270106

Cited by 32 publications

(26 citation statements)

References 18 publications

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“…The main advantage of FIB-SEM imaging is permitting faster visualization of serial ultra-thin sections, which optimizes the adjustments needed for identification of the target structures (23,24). As a result, we confirmed that chondrocytes in the proliferative and the hypertrophic zones extend their short cytoplasmic processes to the cartilaginous fibrils that parallel the longitudinal axis of the growth plate, and that vesicular structures are present in the intercolumnar septa -even at the proliferative zone (Figs.…”

Section: Discussionsupporting

confidence: 70%

Biological application of focus ion beam-scanning electron microscopy (FIB-SEM) to the imaging of cartilaginous fibrils and osteoblastic cytoplasmic processes

Hasegawa

Endo

Tsuchiya

et al. 2017

Journal of Oral Biosciences

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Objectives: The aim of this study is a biological application of focused ion beam-scanning electron microscopy (FIB-SEM) to demonstrate serial sectional images of skeletal tissues, here presenting the ultrastructure of 1) cartilaginous extracellular fibrils and 2) osteoblastic cytoplasmic processes.Methods: Seven weeks-old female wild-type mice were fixed with half-Karnovsky solution and subsequent OsO4, and the tibiae were extracted for block staining prior to observation under transmission electron microscope (TEM) and FIB-SEM.Results: TEM showed the fine fibrillar, but somewhat amorphous ultrastructure of the intercolumnar septa in the growth plate cartilage. Alternatively, FIB-SEM revealed bundles of stout fibrils at regular intervals paralleling the septa's longitudinal axis, as well as vesicular structures embedded in the cartilaginous matrix of the proliferative zone. In the primary trabeculae, both TEM and FIB-SEM showed several osteoblastic cytoplasmic processes on the osteoid, with numbers higher than those seen in the bone matrix. FIB-SEM revealed the agglomeration of cytoplasmic processes beneath the osteoblasts, which formed a tubular continuum extending from those cells. Based on these findings, we postulated that osteoblasts not only extend their cytoplasmic processes through to the bone matrix, but also stack these cell processes on the osteoid of the primary trabeculae. Conclusion:Taken together, it is likely that FIB-SEM imaging strategy on serial sections may successfully deliver new insights on the ultrastructure of cartilage and bone tissues. words

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Section: Discussionsupporting

confidence: 70%

Biological application of focus ion beam-scanning electron microscopy (FIB-SEM) to the imaging of cartilaginous fibrils and osteoblastic cytoplasmic processes

Hasegawa

Endo

Tsuchiya

et al. 2017

Journal of Oral Biosciences

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“…AFM and SEM are appropriate techiques to observe biological material [33,34]. By operating AFM in tapping mode we reduced the friction and adhesive force and enabled imaging of soft biological samples.…”

Section: Discussionmentioning

confidence: 99%

Blood and Synovial Microparticles as Revealed by Atomic Force and Scanning Electron Microscope

Junkar¹,

Šuštar²,

Frank³

et al. 2009

TOAUTOJ

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Abstract:Microvesicles which are pinched off the cell membrane can be considered extracellular organelles which mediate interaction between distal cells. They were suggested to play an important role in many diseases including autoimmune disorders, however, standard methods for their assessment have not yet been decided upon while their clinical relevance and the underlying mechanisms are yet unclear. We present a pilot study results involving atomic force microscope (AFM) and scanning electron microscope (SEM) images of the material isolated from peripheral blood of healthy donors and from synovial fluid of patients with psoriatic arthritis and rheumatoid arthritis, which is expected to contain microvesicles. Micrographs reveal in the samples isolated from blood the presence of globular and tubular structures which are most probably microvesicles while the identity of grain-like structures isolated from synovial fluid remains obscure. To the best of our knowledge the AFM and SEM images of the material isolated from synovial fluid are presented for the first time.

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“…Instruments with this "slice-and-view" capability are available commercially, and have been used extensively to view the internal architecture of semiconductors and other solids (Giannuzzi et al, 2002;Giannuzzi and Stevie, 2005;Sugiyama and Sigesato, 2004). Preliminary efforts of the use of focused ion beam microscopy to view the internal structure of biological specimens have also been reported (Ballerini et al, 2001;Drobne et al, 2005;Gestmann et al, 2004;Heymann et al, 2005;Mulders, 2003;Young et al, 1993). Here, we report on our progress in applying dual beam imaging to biologically relevant samples at room temperature and at cryogenic temperatures.…”

Section: Introductionmentioning

confidence: 99%

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

Heymann

Hayles²,

Gestmann³

et al. 2006

Journal of Structural Biology

318

240

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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.

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Focused ion beam/scanning electron microscopy studies of Porcellio scaber (Isopoda, Crustacea) digestive gland epithelium cells

Cited by 32 publications

References 18 publications

Biological application of focus ion beam-scanning electron microscopy (FIB-SEM) to the imaging of cartilaginous fibrils and osteoblastic cytoplasmic processes

Biological application of focus ion beam-scanning electron microscopy (FIB-SEM) to the imaging of cartilaginous fibrils and osteoblastic cytoplasmic processes

Blood and Synovial Microparticles as Revealed by Atomic Force and Scanning Electron Microscope

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

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