scholarly journals Contrast-Enhanced Tissue Processing of Fibrillin-Rich Elastic Fibres for 3D Visualization by Volume Scanning Electron Microscopy

2021 ◽  
Vol 4 (3) ◽  
pp. 56
Author(s):  
Philip N. Lewis ◽  
Robert D. Young ◽  
R. B. Souza ◽  
Andrew J. Quantock ◽  
Keith M. Meek
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
3D Visualization ◽  
Elastic Fibre ◽  
Microscopical Examination ◽  
Elastic Fibres ◽  
Tissue Samples ◽  
Embedded Blocks ◽  
3D Architecture ◽  
Scanning Electron

Elastic fibres constitute an important component of the extracellular matrix and currently are the subject of intensive study in order to elucidate their assembly, function and involvement in cell–matrix interactions and disease. However, few studies to date have investigated the 3D architecture of the elastic fibre system in bulk tissue. We describe a protocol for preparation of tissue samples, including primary fixation and backscatter electron contrast-enhancement steps, through dehydration into stable resin-embedded blocks for volume electron microscopy. The use of low molecular weight tannic acid and alcoholic lead staining are critical stages in this procedure. Block preparation by ultramicrotomy and evaporative metal coating prior to microscopical examination are also described. We present images acquired from serial block face scanning electron microscopy of cornea and aorta showing target structures clearly differentiated from cells and other matrix components. The processing method imparts high contrast to fibrillin-containing elastic fibres, thus facilitating their segmentation and rendering into 3D reconstructions by image analysis software from large serial image datasets.

Serial Block-Face Scanning Electron Microscopy (SBF-SEM) of Biological Tissue Samples

10.3791/62045 ◽  
2021 ◽  
Author(s):  
Justin A. Courson ◽  
Paul T. Landry ◽  
Thao Do ◽  
Eric Spehlmann ◽  
Pascal J. Lafontant ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Biological Tissue ◽  
Tissue Samples ◽  
Face Scanning ◽  
Block Face ◽  
Scanning Electron

scholarly journals Investigation of the Presence of Biofilms in Chronic Suppurative Otitis Media, Nonsuppurative Otitis Media, and Chronic Otitis Media with Cholesteatoma by Scanning Electron Microscopy

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Ercan Kaya ◽  
Ilknur Dag ◽  
Armagan Incesulu ◽  
Melek Kezban Gurbuz ◽  
Mustafa Acar ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Otitis Media ◽  
Middle Ear ◽  
Chronic Otitis Media ◽  
Chronic Suppurative Otitis Media ◽  
Middle Ear Mucosa ◽  
Suppurative Otitis Media ◽  
Tissue Samples ◽  
Scanning Electron

Objective. Biofilms have been shown to play a major role in the pathogenesis of otolaryngologic infections. However, very limited studies have been undertaken to demonstrate the presence of biofilms in tissues from patients with chronic otitis media (COM) with or without cholesteatoma. Our objective is to study the presence of biofilms in humans with chronic suppurative and nonsuppurative otitis media and cholesteatoma.Study Design. In all, 102 tissue specimens (middle ear, mastoid tissue, and ossicle samples) were collected during surgery from 34 patients.Methods. The samples were processed for the investigation of biofilms by scanning electron microscopy (SEM).Results. Our research supports the hypothesis in which biofilms are involved in chronic suppurative otitis media, cholesteatoma, and, to a lesser degree, chronic nonsuppurative otitis media. There were higher rates in hypertrophic and granulated tissue samples than in normal mucosa. In addition, the presence of biofilms was significantly higher in the middle ear mucosa compared with the mastoid and ossicle samples.Conclusion. In the clinic, the careful use of topical or systemic antimicrobials is essential, and, during surgery, hypertrophic tissue must be carefully removed from normal tissue.

scholarly journals Visualizing the 3D Architecture of Multiple Erythrocytes Infected with Plasmodium at Nanoscale by Focused Ion Beam-Scanning Electron Microscopy

PLoS ONE ◽  
2012 ◽  
Vol 7 (3) ◽  
pp. e33445 ◽  
Author(s):  
Lia Carolina Soares Medeiros ◽  
Wanderley De Souza ◽  
Chengge Jiao ◽  
Hector Barrabin ◽  
Kildare Miranda
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Beam Scanning ◽  
3D Architecture ◽  
Scanning Electron

scholarly journals Gobioides broussonnetii (Gobiidae): a new host forPterobothrium crassicolle (Trypanorhyncha) on Marajó Island, northern Brazil

2013 ◽  
Vol 22 (3) ◽  
pp. 398-401 ◽  
Author(s):  
Marcela Videira ◽  
Michele Velasco ◽  
Lilian Dias ◽  
Patrícia Matos ◽  
Henrique Diniz Ferreira de Almeida ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
New Host ◽  
Tissue Samples ◽  
The North ◽  
Brazilian State ◽  
Marajó Island ◽  
Northern Brazil ◽  
Scanning Electron

In the present study, the cestoid Pterobothrium crassicolle Diesing, 1850 (Trypanorhyncha), was observed parasitizing specimens of the violet goby (Gobioides broussonnetii Lacepède, 1800) collected from the estuary of the Paracauarí River on Marajó Island in the north of the Brazilian state of Pará, between January 2009 and December 2010. Tissue samples were analyzed, which led to identification of blastocysts containing plerocercoid larvae. These larvae were processed for scanning electron microscopy. Sixty G. broussonnetii specimens were dissected, and P. crassicolle was found in 48 (80%) of them. The violet goby,G. broussonnetii, is a new host for P. crassicolle.

Combined high-voltage and scanning electron microscopy on the same brain tissue samples for the study of blood-brain barrier injury

1989 ◽  
Vol 47 ◽  
pp. 986-987
Author(s):  
A.S. Lossinsky ◽  
M. J. Song ◽  
R. Pluta ◽  
R. C. Moretz ◽  
H. M. Wisniewski
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Blood Vessels ◽  
High Voltage ◽  
Inflammatory Cells ◽  
Junctional Complex ◽  
Electron Microscopic ◽  
Tissue Samples ◽  
Junctional Complexes ◽  
Scanning Electron

The application of combined techniques utilizing both scanning electron microscopy (SEM) with conventional transmission electron microscopy (CTEM) has a decided advantage because it permits analysis of identical tissue areas. If one is interested in analyzing of endothelial cell (EC) tight junctions, for example, although SEM offers unlimited topographical information, it is of critical importance to concomitantly assess EC junctions by CTEM from adjacent tissue blocks in order to visualize whether or not the junctional complexes are open only at the surface, for example, while deeper regions of this junctional complex may actually be occluded. Generally, three dimensional reconstruction is required employing either serial thin section analysis or high-voltage electron microscopic (HVEM) analysis of thick (0.25-1.0 μm) sections in order to obtain this information.Micro blood vessels (veins and venules) from the thalamus of SJL mice subjected to chronic relapsing experimental allergic encephalomyelitis were used in the present studies for examining EC junctional complexes and perivascular infiltration of inflammatory cells. Animals were perfused, fixed with standards buffered aldehydes, prepared for SEM, and blood vessels were identified and analyzed with an ISI-40 SEM. The same tissue blocks were trimmed to the targeted micro blood vessels, rinsed three times in propylene oxide (p.o.) for 20 min. each and flat-embedded in Spurr plastic following 1 hr. in 1:1 p.o.:Spurr. Thick (0.5 μm) sections were collected on Formvar-coated slot grids, stained with uranyl and lead salts and examined with an AEl HVEM (Albany, NY) at 1000 KV.

Root cementum ultrastructure in healthy and periodontally diseased teeth

2020 ◽  
Vol 25 (4) ◽  
pp. 317-321
Author(s):  
E. S. Slazhneva ◽  
E. A. Tikhomirova ◽  
L. A. Elizova ◽  
E. S. Loboda ◽  
L. Yu. Orekhova ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Significant Role ◽  
Chronic Periodontitis ◽  
Structural Changes ◽  
3D Visualization ◽  
Periodontal Regeneration ◽  
Sem Images ◽  
Dentinal Tubules ◽  
Scanning Electron

Relevance. Investigation of the root cementum ultrastructure in chronic generalized periodontitis is still relevant as changes in structure and composition of root cementum play a significant role in successful periodontal regeneration. Am is to study changes in the root cementum ultrastructure in patients with chronic generalized periodontitis.Materials and methods. Scanning electron microscopy (SEM) was used to study the cementum surface of 9 teeth extracted due to severe chronic generalized periodontitis and 3 teeth with a clinically healthy periodontium extracted for orthodontic reasons. 3D visualization of the received SEM images was performed.Results. The cementum of periodontally healthy teeth appeared homogeneous and regular,was covered in periodontal fibers and had a pebble-like or dome-shaped surface. In chronic periodontitis patients, the cementum surface was mostly irregular with multiple defects of various depth, areas of completely destroyed cementum, exposed dentinal tubules and a complete absence of periodontal fibers.Conclusion. Loss of periodontal attachment and root cementum exposure to microbial biofilm may result in irreversible structural changes of the surface which may affect the regeneration of clinical attachment.

Spatial distribution of organelles in leaf cells and soybean root nodules revealed by focused ion beam-scanning electron microscopy

2018 ◽  
Vol 45 (2) ◽  
pp. 180 ◽  
Author(s):  
Brandon C. Reagan ◽  
Paul J. -Y. Kim ◽  
Preston D. Perry ◽  
John R. Dunlap ◽  
Tessa M. Burch-Smith
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Focused Ion Beam ◽  
Ion Beam ◽  
Plant Tissues ◽  
3D Analysis ◽  
Beam Scanning ◽  
Tissue Samples ◽  
Thin Sections ◽  
Scanning Electron

Analysis of cellular ultrastructure has been dominated by transmission electron microscopy (TEM), so images collected by this technique have shaped our current understanding of cellular structure. More recently, three-dimensional (3D) analysis of organelle structures has typically been conducted using TEM tomography. However, TEM tomography application is limited by sample thickness. Focused ion beam-scanning electron microscopy (FIB-SEM) uses a dual beam system to perform serial sectioning and imaging of a sample. Thus FIB-SEM is an excellent alternative to TEM tomography and serial section TEM tomography. Animal tissue samples have been more intensively investigated by this technique than plant tissues. Here, we show that FIB-SEM can be used to study the 3D ultrastructure of plant tissues in samples previously prepared for TEM via commonly used fixation and embedding protocols. Reconstruction of FIB-SEM sections revealed ultra-structural details of the plant tissues examined. We observed that organelles packed tightly together in Nicotiana benthamiana Domin leaf cells may form membrane contacts. 3D models of soybean nodule cells suggest that the bacteroids in infected cells are contained within one large membrane-bound structure and not the many individual symbiosomes that TEM thin-sections suggest. We consider the implications of these organelle arrangements for intercellular signalling.

Multilayer Microstructure of Idiopathic Epiretinal Macular Membranes

2017 ◽  
Vol 27 (6) ◽  
pp. 762-768 ◽  
Author(s):  
Claudio Azzolini ◽  
Terenzio Congiu ◽  
Simone Donati ◽  
Alberto Passi ◽  
Petra Basso ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Soft Tissue ◽  
Fibrous Material ◽  
Collagen Fibrils ◽  
Thin Sheets ◽  
Inner Limiting Membrane ◽  
3D Architecture ◽  
Fine Resolution ◽  
Scanning Electron

Purpose To identify the ultramicroscopic structure of idiopathic epiretinal macular membranes (iEMMs) by scanning electron microscopy (SEM). Methods We examined 28 iEMMs surgically removed from 28 eyes of 28 patients. All specimens, previously observed at stereomicroscope, were treated with an osmium maceration technique. Fine resolution of iEMMs’ 3D architecture and their interaction with the retina were studied using a Philips SEM-FEG XL-30 microscope. Results The specimens appeared as laminar connective structures partially or completely adherent to the inner limiting membrane (ILM). We identified 4 types of structures: ( 1 ) distinct layers of thin sheets of woven fibers; ( 2 ) folded layers of inhomogeneous thickness of fibrous material more consistent; ( 3 ) thicker and more rigid layers recognizable as collagen fibrils with typical 64-nm period, collagen fibrils isolated or intermingled between them; ( 4 ) lacunar structures with inflammatory and/or necrotic material. The first 3 types of structures appear to thicken towards a centripetal direction from the ILM to the vitreous in order from 1 to 3. The interface of ILM-iEMM tissue shows particular small bridges of connection. Cells are rarely found, especially in the tissue near the ILM. Conclusions Layers of various materials follow one another in iEMMs. Cells are rarely found. The interface ILM-iEMM tissue shows particular small bridges of connection. The dynamic modeling of bended layers begins in soft tissue.

Collagen-calcium phosphate composites

1998 ◽  
Vol 212 (6) ◽  
pp. 413-425 ◽  
Author(s):  
A C Lawson ◽  
J T Czernuszka
Keyword(s):  
Electron Microscopy ◽  
Mechanical Properties ◽  
Scanning Electron Microscopy ◽  
Calcium Phosphate ◽  
Calcium Phosphates ◽  
Hierarchical Architecture ◽  
Microscopical Examination ◽  
Bone Augmentation ◽  
Solid Phases ◽  
Scanning Electron

There is a widespread clinical need for bone augmentation and replacement. The major solid phases of bone are collagen and calcium phosphate and a bone analogue based on these two constituents should have some useful properties. In this review this theme is developed and the properties of natural and naturally based composites are compared. Composites have been produced by the precipitation of calcium phosphates on to collagen and a summary of the methods and results from mechanical testing and scanning electron microscopy are presented. Composites with mechanical properties intermediate between cancellous and cortical bone have been produced. The review concludes by explaining some of the mechanical properties of the composites, using knowledge of the hierarchical architecture of bone and results from microscopical examination of the fractured composites.

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