Confocal laser scanning microscopy (CLSM) was used to examine archaeoparasitological specimens from coprolites associated with La Cueva de los Muertos Chiquitos (CMC) located near present-day Durango, Mexico. The eggs for 4 different types of parasites recovered from CMC coprolites were imaged using CLSM to assist with identification efforts. While some of the parasite eggs recovered from CMC coprolites were readily identified using standard light microscopy (LM), CLSM provided useful data for more challenging identifications by highlighting subtle morphological features and enhancing visualization of parasite egg anatomy. While other advanced microscopy techniques, such as scanning electron microscopy (SEM), may also detect cryptic identifying characters, CLSM is less destructive to the specimens. Utilizing CLSM allows for subsequent examinations, such as molecular analyses, that cannot be performed following SEM sample preparation and imaging. Furthermore, CLSM detects intrinsic autofluorescence molecules, making improved identification independent of resource and time-intensive protocols. These aspects of CLSM make it an excellent method for assisting in taxonomic identification and for acquiring more detailed images of archaeoparasitological specimens.
Eggs ; Methods ; Mexico ; Microscopy ; Microscopy, Confocal ; Microscopy, Electron, Scanning ; Ovum ; Parasites

OBJECTIVETo provide practical method for microscopic authentication of traditional Chinese medicine Gusuibu and its adulterants.

METHODBy means of light microscope, scanning electron microscopy and tissue section techniques, the morphology, the size of the rhizome scales and their bearing position in the original plants of Gusuibu and its adulterants, i. e. Drynaria roosii, D. delavayi, D. quercifolia and Pseudodrynaria coronans were analyzed.

RESULTThere were significant differences between scales length of D. roosii, D. delavayi and P. coronans, while there was no significant difference between that of D. roosii and D. quercifolia. The scale teeth of D. delavayi were usually curved, bifid and uneven distributed at the scale fringe, which was different from that of the other three species. The base of the scales sinks in epidermis in D. roosii, D. quercifolia, and P. coronans, while it bore at the raised part of epidermis in D. delavayi.

CONCLUSION[corrected] Morphology, size and bearing position of the rhizome scales have significant differences in the several species. Therefore, these characteristics can be applied to the identification of Gusuibu and its adulterants.

Medicine, Chinese Traditional ; methods ; Microscopy ; Microscopy, Electron, Scanning ; Polypodiaceae ; anatomy & histology ; classification ; Rhizome ; anatomy & histology ; classification

OBJECTIVETo compare the efficacy of three methods for forensic diatom test, namely strong acid digestion-centrifuge enrichment-light microscopy (SD-CE-LM), microwave digestion-membrane filtration-automated scanning electron microscopy (MD-ME-SEM), and microwave digestion-membrane filtration-light microscopy (MD-MF-LM).

METHODSSixty samples were randomly divided into 3 groups for diatom test using three methods, and the sample preparation time, degree of digestion and recovery rate of diatoms were compared.

RESULTSThe sample preparation time was the shortest with MD-MF-LM and the longest with SD-CE-LM (P<0.05). MD-ME-SEM and MD-MF-LM allowed more thorough tissue digestion than SD-CE-LM. MD-ME-SEM resulted in the highest total recovery rate of diatom, followed by MD-MF-LM and then by SD-CE-LM (P<0.05); the recover rate of different diatom species was the highest with MD-ME-SEM, followed by MD-MF-LM and SD-CE-LM (P<0.05).

CONCLUSIONSD-CE-LM has a low recovery rate of diatoms especially for those with lengths shorter than 40 µm or densities less than 1/5. With a high recovery rate and accuracy in diatom test, MD-ME-SEM is suitable for diagnosis of suspected drowning cases. MD-MF-LM is highly efficient, sensitive and convenient for forensic diatom test.

Centrifugation ; Diatoms ; isolation & purification ; Drowning ; Forensic Sciences ; methods ; Humans ; Microscopy ; Microscopy, Electron, Scanning ; Microwaves ; Specimen Handling

OBJECTIVETo study the histology and ultrastructure of the tidemark in the adult condylar cartilage and their significance.

METHODSAfter embedded in paraffin, 50 adult condyles were stained with HE, partly with Van-Gieson and histochemical methods, then observed by light microscope. 3 cases of the tidemark region were observed by transmission electron microscope, another 5 cases were studied by scanning electron microscope.

RESULTSIn the tidemark region, there had shown the presence of the AKP and calcium, absence of the proteoglycan, abundance of the membrane-bound matrix vesicles, crystals of hydroxyapatite and lipid nodule-like substances, which were often observed in the load-bearing areas. The collagen fibrils of the noncalcified cartilage crossed the tidemark gradiently and were continuous with those of the calcified cartilage; a band of horizontal fibrils surrounded the whole tidemark region, which was wider in the load-bearing areas than that in the nonload-bearing areas and which interweaved with the gradient fibrils so as to form a net. Digested with papain, the surface of the tidemark was highly undulating, and a lot of chondrocyte lacunae were seen on the surface, which were surrounded by calcified tissues.

CONCLUSIONSIn the region of the tidemark, physiological calcification takes place and is more active in the load-bearing areas; gradient and horizontal fibrils interweave with each other, which is correlated with the force on the articulation.

Adult ; Female ; Histocytochemistry ; methods ; Humans ; Male ; Mandibular Condyle ; cytology ; ultrastructure ; Microscopy, Electron ; Microscopy, Electron, Scanning ; Temporomandibular Joint ; cytology ; ultrastructure

Vitrification is a promising alternative to tissue preservation, which will greatly avoid the ice-crystal formation and circumvent the hazardous effects associated with ice formation during the entire procedures. In this study, we evaluate the effect of vitreous cryopreservation of rabbit trachea by comparing the vitrification procedure with the conventional computer-programmed slow freezing approaches. Harvested tracheae were tailored and divided into groups and cryopreserved by vitrification and programmed freezing, respectively. The morphology and ultrastructure of the thawed tracheal fragments were examined by using HE staining, TUNEL test, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) to assess the integrity of the tracheal fragments. The morphological studies demonstrated both cryopreservation procedure retain the interity of trachea, and that epithelium mucosae, cilia and cartilage cells were in good shape. Compared with slow freezing methods, vitrification was less detrimental to cartilage cells and had a higher survival rate of chondrocytes and coverage of epithelium and cilia. Therefore, vitrification method is a more satisfactory method to preserve trachea, the survival of chondrocytes in situ in cartilage tissue is adequate, and respiratory epithelium is soundly preserved.
Animals ; Cryopreservation ; methods ; Cryoprotective Agents ; chemistry ; Microscopy, Electron, Scanning ; Rabbits ; Tissue Preservation ; methods ; Trachea ; cytology ; ultrastructure

p-Nitrophenylphosphate (PNPP) is usually employed as the substrate for enzyme-linked immunosorbent assays. p-Nitrophenol (PNP), the product of PNPP, with the catalyst alkaline phosphatase (ALP), will passivate an electrode, which limits applications in electrochemical analysis. A novel anti-passivation ink used in the preparation of a graphene/ionic liquid/chitosan composited (rGO/IL/Chi) electrode is proposed to solve the problem. The anti-passivation electrode was fabricated by directly writing the graphene-ionic liquid-chitosan composite on a single-side conductive gold strip. A glassy carbon electrode, a screen-printed electrode, and a graphene-chitosan composite-modified screen-printed electrode were investigated for comparison. Scanning electron microscopy was used to characterize the surface structure of the four different electrodes and cyclic voltammetry was carried out to compare their performance. The results showed that the rGO/IL/Chi electrode had the best performance according to its low peak potential and large peak current. Amperometric responses of the different electrodes to PNP proved that only the rGO/IL/Chi electrode was capable of anti-passivation. The detection of cardiac troponin I was used as a test example for electrochemical immunoassay. Differential pulse voltammetry was performed to detect cardiac troponin I and obtain a calibration curve. The limit of detection was 0.05 ng/ml.
Electrochemical Techniques/methods* ; Electrodes ; Graphite ; Immunoassay/methods* ; Ink ; Microscopy, Electron, Scanning ; Troponin I/blood*

Clove oil and turmeric oil were absorbed by mesoporous carbon. The absorption ratio of mesoporous carbon to volatile oil was optimized with the eugenol yield and curcumol yield as criteria Curing powder was characterized by scanning electron microscopy (SEM) and differential scanning calorietry (DSC). The effects of mesoporous carbon on dissolution in vitro and thermal stability of active components were studied. They reached high adsorption rate when the absorption ratio of mesoporous carbon to volatile oil was 1:1. When volatile oil was absorbed, dissolution rate of active components had a little improvement and their thermal stability improved after volatile oil was absorbed by the loss rate decreasing more than 50%. Absorbing herbal volatile oil with mesoporous carbon deserves further studying.
Adsorption ; Carbon ; chemistry ; Drug Stability ; Microscopy, Electron, Scanning ; methods ; Oils, Volatile ; chemistry ; Sesquiterpenes ; chemistry

Conventional sample preparation technique for adipose tissue is difficult to achieve satisfactory results for scanning electron microscopy (SEM). We adopted a strategy of postfix with osmium tetroxide to stabilize the fatty acids, phospholipids protein, and hence the membrane structure. Also by extending the dehydration time to fully replace the organic solvents, we achieved satisfactory results for SEM of adipose samples.
Adipose Tissue ; ultrastructure ; Animals ; Female ; Humans ; Microscopy, Electron, Scanning ; Rats ; Rats, Sprague-Dawley ; Specimen Handling ; methods

PURPOSE: The object of the present study was to evaluate the shear bonding strength of composite to PEKK by applying several methods of surface treatment associated with various bonding materials. MATERIALS AND METHODS: One hundred and fifty PEKK specimens were assigned randomly to fifteen groups (n = 10) with the combination of three different surface treatments (95% sulfuric acid etching, airborne abrasion with 50 µm alumina, and airborne abrasion with 110 µm silica-coating alumina) and five different bonding materials (Luxatemp Glaze & Bond, Visio.link, All-Bond Universal, Single Bond Universal, and Monobond Plus with Heliobond). After surface treatment, surface roughness and contact angles were examined. Topography modifications after surface treatment were assessed with scanning electron microscopy. Resin composite was mounted on each specimen and then subjected to shear bond strength (SBS) test. SBS data were analyzed statistically using two-way ANOVA, and post-hoc Tukey's test (P<.05). RESULTS: Regardless of bonding materials, mechanical surface treatment groups yielded significantly higher shear bonding strength values than chemical surface treatment groups. Unlike other adhesives, MDP and silane containing self-etching universal adhesive (Single Bond Universal) showed an effective shear bonding strength regardless of surface treatment method. CONCLUSION: Mechanical surface treatment behaves better in terms of PEKK bonding. In addition, self-etching universal adhesive (Single Bond Universal) can be an alternative bonding material to PEKK irrespective of surface treatment method.
Adhesives ; Aluminum Oxide ; Dental Bonding ; Methods ; Microscopy, Electron, Scanning ; Polymers* ; Sulfur

In the present study, we aimed to determine the cause of surface film formation in three rice vinegars fermented using the traditional static fermentation method. The pH and total acidity of vinegar were 3.0–3.3 and 3.0–8.7%, respectively, and acetic acid was the predominant organic acid present. Colonies showing a clear halo on GYC medium were isolated from the surface film of all vinegars. Via 16S rDNA sequencing, all of the isolates were identified as Acetobacter pasteurianus. Furthermore, field-emission scanning electron microscopy analysis showed that the bacterial cells had a rough surface, were rod-shaped, and were ∼1 × 2 µm in size. Interestingly, cells of the isolate from one of the vinegars were surrounded with an extremely fine threadlike structure. Thus, our results suggest that formation of the surface film in rice vinegar was attributable not to external contamination, to the production of bacterial cellulose by A. pasteurianus to withstand the high concentrations of acetic acid generated during fermentation. However, because of the formation of a surface film in vinegar is undesirable from an industrial perspective, further studies should focus on devising a modified fermentation process to prevent surface film formation and consequent quality degradation.
Acetic Acid ; Acetobacter ; Cellulose ; DNA, Ribosomal ; Fermentation ; Hydrogen-Ion Concentration ; Methods ; Microscopy, Electron, Scanning