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 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 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 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*

PURPOSE: The aim of this study was to test the modulus of elasticity (E) across the interfaces of yttria stabilized zirconia (YTZP) / veneer multilayers using nanoindentation. MATERIALS AND METHODS: YTZP core material (KaVo-Everest, Germany) specimens were either coated with a liner (IPS e.max ZirLiner, Ivoclar-Vivadent) (Type-1) or left as-sintered (Type-2) and subsequently veneered with a pressable glass-ceramic (IPS e.max ZirPress, Ivoclar-Vivadent). A 5 µm (nominal tip diameter) spherical indenter was used with a UMIS CSIRO 2000 (ASI, Canberra, Australia) nanoindenter system to test E across the exposed and polished interfaces of both specimen types. The multiple point load – partial unload method was used for E determination. All materials used were characterized using Scanning Electron Microscopy (SEM) and X – ray powder diffraction (XRD). E mappings of the areas tested were produced from the nanoindentation data. RESULTS: A significantly (P<.05) lower E value between Type-1 and Type-2 specimens at a distance of 40 µm in the veneer material was associated with the liner. XRD and SEM characterization of the zirconia sample showed a fine grained bulk tetragonal phase. IPS e-max ZirPress and IPS e-max ZirLiner materials were characterized as amorphous. CONCLUSION: The liner between the YTZP core and the heat pressed veneer may act as a weak link in this dental multilayer due to its significantly (P<.05) lower E. The present study has shown nanoindentation using spherical indentation and the multiple point load - partial unload method to be reliable predictors of E and useful evaluation tools for layered dental ceramic interfaces.
Ceramics* ; Elastic Modulus ; Hot Temperature ; Methods ; Microscopy, Electron, Scanning ; Powder Diffraction

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

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

BACKGROUNDCurrently, several systems of dentin substrate-reacting adhesives are available for use in the restorative treatment against caries. However, the bond effectiveness and property of different adhesive systems to caries-affected dentin are not fully understood. The objective of this study was to evaluate the bond strength of different adhesives to both normal dentin (ND) and caries-affected dentin (CAD) and to analyze the dentin/adhesive interfacial characteristics.

METHODSTwenty eight extracted human molars with coronal medium carious lesions were randomly assigned to four groups according to adhesives used. ND and CAD were bonded with etch-and-rinse adhesive Adper Single Bond 2 (SB2) or self-etching adhesives Clearfil SE Bond (CSE), Clearfil S(3) Bond (CS3), iBond GI (IB). Rectangular sticks of resin-dentin bonded interfaces 0.9 mm(2) were obtained. The specimens were subjected to microtensile bond strength (microTBS) testing at a crosshead speed of 1 mm/min. Mean microTBS was statistically analyzed with analysis of variance (ANOVA) and Student-Newman-Keuls tests. Interfacial morphologies were analyzed by Scanning Electron Microscopy (SEM).

RESULTSEtch-and-rinse adhesive Adper(TM) Single Bond 2 yielded high bond strength when applied to both normal and caries-affected dentin. The two-step self-etching adhesive Clearfil SE Bond generated the highest bond strength to ND among all adhesives tested but a significantly reduced strength when applied to CAD. For the one-step self-etching adhesives, Clearfil S(3) Bond and iBond GI, the bond strength was relatively low regardless of the dentin type. SEM interfacial analysis revealed that hybrid layers were thicker with poorer resin tag formation and less resin-filled lateral branches in the CAD than in the ND for all the adhesives tested.

CONCLUSIONThe etch-and-rinse adhesive performed more effectively to caries-affected dentin than the self-etching adhesives.

Adhesives ; Dental Bonding ; methods ; Dentin ; Humans ; Microscopy, Electron, Scanning ; Molar ; Tensile Strength