scholarly journals Evaluation of Dental Composite Resin Interface and Adhesion Using Laser Scanning Confocal Microscopy

2018 ◽  
Vol 55 (4) ◽  
pp. 541-544
Author(s):  
Adrian Tudor Stan ◽  
Adrian Vaduva ◽  
Meda Lavinia Negrutiu ◽  
Cosmin Sinescu ◽  
Camelia Szuhanek ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Composite Resin ◽  
Laser Scanning Confocal Microscopy ◽  
Composite Resins ◽  
Dental Composite ◽  
Background Information ◽  
Confocal Laser ◽  
Scanning Confocal Microscopy ◽  
Dental Composite Resin

The aim of the study was to evaluate and to compare the interface and marginal leakage of dental composite resins using confocal laser scanning biological microscopy (CM). Ten class II cavities were prepared on the proximal surfaces of 10 extracted teeth, previously kept in 10% formalin to preserve the bonding properties of the tooth structures. Cavities were restored with dental composite resin using the �open sandwich� technique and then assessed using CM. Open marginal adaptations at the interfaces inside the composite resins were identified. The results obtained with CM helped to eliminate and reduce the background information collected from a distance from the focal plane and allowed for the acquisition of serial optical sections from thick tooth specimens. In vitro assessments were thus made using CM on the teeth structure, regarding the occurrence of potential micro-leakages at the restoration interface.

Synthesis, singlet oxygen generation, photocytotoxicity and subcellular localization of azobisporphyrins as potentially photodynamic therapeutic agents in vitro cell study

2017 ◽  
Vol 21 (02) ◽  
pp. 122-127 ◽  
Author(s):  
Yunman Zheng ◽  
Sizhe Zhu ◽  
Lijun Jiang ◽  
Fengshou Wu ◽  
Chi Huang ◽  
...  
Keyword(s):  
Singlet Oxygen ◽  
Hela Cells ◽  
Cellular Localization ◽  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Confocal Laser Scanning Microscope ◽  
Confocal Laser ◽  
Oxygen Generation ◽  
Scanning Confocal Microscopy

Three azobisporphyrins (Por1, Por2 and Por3) were synthesized by coupling two molecules of (4-nitrophenyl/pyridyl) porphyrins in the presence of KOH/butanol. The structures of porphyrins were confirmed by UV, IR, NMR and mass spectra and elemental analysis. With tetraphenylporphyrin (H2TPP) as a control, the singlet oxygen (1O[Formula: see text] generation of porphyrins was evaluated through 1,3-diphenylisobenzofuran (DPBF) method. The order of ability to generate 1O2 for three azobisporphyrins was Por 1 [Formula: see text]Por 2 > Por 3[Formula: see text] H2TPP. The photocytotoxicity and sub-cellular localization of azobisporphyrins over Hela cells were studied through MTT analysis and confocal laser scanning microscope, respectively. The results indicated Por 1 and Por 2 displayed the low dark-cytotoxicity, while Por 3 induced a concentration-dependent cytotoxicity to Hela cells with the concentration of porphyrins ranging from 1 to 100 [Formula: see text] M. With the light dose at 4 J/cm2, Por 3 killed more than 60% Hela cells at 2 [Formula: see text] M, indicating a high photocytoxicity. As seen from the laser scanning confocal microscopy images, Por 3 was mainly localized in cell membrane, while Por 1 and Por 2 do not displayed significant fluorescent emission in Hela cells. These results suggest the synthesized cationic azobisporphyrin could be used as a potential therapeutic agent for photodynamic therapy of cancers.

scholarly journals An In Vitro Comparative Evaluation of the Effects of Different Bleaching Agents on Marginal Integrity of Two Different Restorative Resin Materials: An Analysis by Confocal Laser Microscopy

2019 ◽  
Vol 07 (03) ◽  
pp. 114-122
Author(s):  
Nitesh Goyal ◽  
Yogesh Kumar ◽  
Neetu Jindal ◽  
Renu Aggarwal
Keyword(s):  
Laser Scanning ◽  
Composite Resin ◽  
Root Surface ◽  
Composite Resins ◽  
Tooth Bleaching ◽  
Confocal Laser ◽  
Control Group ◽  
Class V ◽  
Nanohybrid Composite

Abstract Introduction In our society, with the advances in cosmetic consciousness, well-aligned, properly contoured, white and clean teeth are a symbol of health and standard of beauty. Thus, tooth bleaching has become popular cosmetic therapy among patients and dentists. Microleakage is a major challenge to the success rate for all type of restorations. Materials and Methods Eighty maxillary human molars were collected and evaluated and Class V cavities 5×3×2 mm size were prepared with straight carbide bur (SS White) on the buccal and palatal surfaces at the cementoenamel junction of all the samples, so that the upper margin would be in enamel and lower margins on root surface. In total, 160 class V cavities were prepared on maxillary molars. Then samples were randomly divided into 3 groups (microfilled and nanohybrid composite resins and control group) and 3 subgroups (Pola Office, Whiteness HP Blue, Whiteness Perfect) according to the type of composite resins and bleaching agents used. After application of bleaching agents, all samples were immersed in a contrast solution of rhodamine B fluorescent dye for 24 hours. Then using a diamond disc samples were sectioned buccolingually through the center of the restorations and sectioned teeth were evaluated for marginal microleakage under confocal laser scanning microscope to check the extent of dye penetration. Results When all composite resin groups were compared with all the bleaching agents, at occlusal and gingival levels, lowest marginal leakage scores were obtained with the microfilled composite resin bleached with Whiteness HP blue, and in nanohybrid composite lowest microleakage score were obtained with Pola office bleach at both levels.

scholarly journals Advanced Static and Dynamic Fluorescence Microscopy Techniques to Investigate Drug Delivery Systems

Pharmaceutics ◽  
2021 ◽  
Vol 13 (6) ◽  
pp. 861
Author(s):  
Jacopo Cardellini ◽  
Arianna Balestri ◽  
Costanza Montis ◽  
Debora Berti
Keyword(s):  
Drug Delivery ◽  
Fluorescence Microscopy ◽  
Drug Delivery Systems ◽  
Laser Scanning ◽  
Super Resolution ◽  
Laser Scanning Confocal Microscopy ◽  
Delivery Systems ◽  
Scanning Confocal Microscopy ◽  
Biological Phenomena

In the past decade(s), fluorescence microscopy and laser scanning confocal microscopy (LSCM) have been widely employed to investigate biological and biomimetic systems for pharmaceutical applications, to determine the localization of drugs in tissues or entire organisms or the extent of their cellular uptake (in vitro). However, the diffraction limit of light, which limits the resolution to hundreds of nanometers, has for long time restricted the extent and quality of information and insight achievable through these techniques. The advent of super-resolution microscopic techniques, recognized with the 2014 Nobel prize in Chemistry, revolutionized the field thanks to the possibility to achieve nanometric resolution, i.e., the typical scale length of chemical and biological phenomena. Since then, fluorescence microscopy-related techniques have acquired renewed interest for the scientific community, both from the perspective of instrument/techniques development and from the perspective of the advanced scientific applications. In this contribution we will review the application of these techniques to the field of drug delivery, discussing how the latest advancements of static and dynamic methodologies have tremendously expanded the experimental opportunities for the characterization of drug delivery systems and for the understanding of their behaviour in biologically relevant environments.

scholarly journals 273 OPTIMIZATION OF PROTOCOLS FOR ACTIVATION OF GOAT OOCYTES WITH IONOMYCIN IN COMBINATION WITH 6-DIMETHYLAMINOPURINE

2005 ◽  
Vol 17 (2) ◽  
pp. 286
Author(s):  
J.-H. Tan ◽  
G.-C. Lan ◽  
Z.-L. Chang ◽  
D. Han ◽  
Z.-B. Han ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Cumulus Cell ◽  
Laser Scanning Confocal Microscopy ◽  
Oocyte Activation ◽  
Scanning Confocal Microscopy ◽  
Multiple Comparison Test ◽  
Parthenogenetic Development ◽  
Activation Rate ◽  
Activation Rates

The protocol of ionomycin in combination with 6-dimethylaminopurine (6-DMAP) is commonly used for activation of oocytes and reconstructed embryos of different species. Since numerous abnormalities and impaired development have been observed when oocytes are activated with 6-DMAP, this protocol needs optimization. Effects of concentration and treatment duration of both drugs on activation kinetics and parthenogenetic development of goat oocytes were examined in this study. When goat oocytes matured in vitro in TCM-199 were treated with 5 M ionomycin in PBS for different periods before exposure to 6-DMAP in CR1aa, the activation rate obtained with ionomycin treatment for 1 min (95.2%) was significantly (P < 0.05, Duncan multiple comparison test) higher than with ionomycin treatments for 3, 5, 7, or 9 min. When oocytes were treated with different concentrations of ionomycin for 1 min before exposure to 6-DMAP, activation rates obtained with 0.625, 1.25, 2.5, 5, 10, and 20 M ionomycin (87–95%) did not differ significantly but were significantly higher than that achieved with 0.3125 M ionomycin. Progressive reduction of time for 6-DMAP exposure showed that the duration of 6-DMAP treatment can be reduced to 1 h from the 2nd up to the 4th hour after ionomycin treatment, to produce activation rates greater than 85%. When oocytes were treated with different concentrations of 6-DMAP for the 3rd hour (atotal of 1 h, 3 h after the exposure to ionomycin), activation rates with 4 and 2 mM 6-DMAP (>90%) were significantly higher than those with 1and 0.5 mM. Therefore, the best protocol for goat oocyte activation would be a 1-min exposure to 2.5 M ionomycin followed by 2 mM 6-DMAP treatment for the 3rd hour. When oocytes matured in vitro for different times were stimulated with the best protocol, activation rates of the 27-, 30-, and 33-h oocytes (85, 85, and 91%) were significantly higher than those of the 24-, 26-, and 39-h oocytes. When activated oocytes were co-cultured in CR1aa with cumulus cell monolayers, the highest rates of cleavage (92%) and morulae/blastocysts (23%) were obtained with oocytes activated by the best protocol, and any increase in the intensity of ionomycin treatment and in the duration of 6-DMAP exposure impaired the development of the parthenotes. During anaphase II, chromosomes (the dyads) did not separate into two units in oocytes that were activated by long exposure to 6-DMAP, but they did in oocytes that were activated by short or no exposure to 6-DMAP; as a result, only one pronucleus developed in most of the former but two pronuclei were formed in most of the latter cases. Laser scanning confocal microscopy showed that microtubules also behaved differently in these two groups of activated oocytes. It is therefore concluded that to obtain better activation and development, goat oocytes matured in vitro for 27–33 h should be used, and these should be activated by a 1-min exposure to 2.5 M ionomycin followed by 2 mM 6-DMAP treatment for the 3rd hour. This study was supported by the “973” Project of China Sci. Technol. Ministry (No. G200016108).

scholarly journals Monitoring Tritrophic Biocontrol Interactions Between Bacillus spp., Fusarium oxysporum f. sp. cubense, Tropical Race 4, and Banana Plants in vivo Based on Fluorescent Transformation System

2021 ◽  
Vol 12 ◽  
Author(s):  
Ping He ◽  
Shu Li ◽  
Shengtao Xu ◽  
Huacai Fan ◽  
Yongfen Wang ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Fluorescent Protein ◽  
Laser Scanning Confocal Microscopy ◽  
Transformation System ◽  
Scanning Confocal Microscopy ◽  
Bacillus Spp ◽  
Green Fluorescent ◽  
Race 4

Bacillus spp. is effective biocontrol agents for Fusarium wilt of banana (FWB), tropical race 4 (TR4). This study explores the colonization by Bacillus subtilis, Bacillus velezensis, and Bacillus amyloliquefaciens of host banana plants and elucidates the mechanism of antagonistic TR4 biocontrol. The authors selected one B. subtilis strain, three B. velezensis strains, and three B. amyloliquefaciens strains that are proven to significantly inhibit TR4 in vitro, optimized the genetic transformation conditions and explored their colonization process in banana plants. The results showed that we successfully constructed an optimized fluorescent electro-transformation system (OD600 of bacteria concentration=0.7, plasmid concentration=50ng/μl, plasmid volume=2μl, transformation voltage=1.8kV, and transformation capacitance=400Ω) of TR4-inhibitory Bacillus spp. strains. The red fluorescent protein (RFP)-labeled strains were shown to have high stability with a plasmid-retention frequency above 98%, where bacterial growth rates and TR4 inhibition are unaffected by fluorescent plasmid insertion. In vivo colonizing observation by Laser Scanning Confocal Microscopy (LSCM) and Scanning Electron Microscopy (SEM) showed that Bacillus spp. can colonize the internal cells of banana plantlets roots. Further, fluorescent observation by LSCM showed these RFP-labeled bacteria exhibit chemotaxis (chemotaxis ratio was 1.85±0.04) toward green fluorescent protein (GFP)-labeled TR4 hyphae in banana plants. We conclude that B. subtilis, B. velezensis, and B. amyloliquefaciens can successfully colonize banana plants and interact with TR4. Monitoring its dynamic interaction with TR4 and its biocontrol mechanism is under further study.

Distribution of a nuclear envelope antigen during the syncytial mitoses of the early Drosophila embryo as revealed by laser scanning confocal microscopy

1992 ◽  
Vol 102 (2) ◽  
pp. 299-305
Author(s):  
M.G. Riparbelli ◽  
G. Callaini
Keyword(s):  
Nuclear Envelope ◽  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Confocal Laser Scanning Microscope ◽  
Drosophila Embryo ◽  
Confocal Laser ◽  
Cytoplasmic Staining ◽  
Scanning Confocal Microscopy ◽  
Early Drosophila Embryo ◽  
Envelope Antigen

The changing distribution of a nuclear envelope antigen recognized by a monoclonal antibody raised against human fibroblast vimentin during the syncytial mitoses of the Drosophila embryo has been studied with a confocal laser scanning microscope. The antigen appears very early as irregular aggregates in the peripheral cytoplasm of the preblastoderm embryo. As the first nuclei reach the periplasm the antigen is localized on the nuclear envelope and the cytoplasmic staining decreases. In addition to the perinuclear labeling we observed intense midzone and polar staining during the mitotic cycle. A possible relationship between polar localization of the antigen and centrosome position is discussed.

Initiation of appressorium formation in Uromyces appendiculatus: organization of the apex, and the responses involving microtubules and apical vesicles

1991 ◽  
Vol 69 (11) ◽  
pp. 2560-2573 ◽  
Author(s):  
Young H. Kwon ◽  
Harvey C. Hoch ◽  
James R. Aist
Keyword(s):  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Uromyces Appendiculatus ◽  
Signal Recognition ◽  
Appressorium Formation ◽  
Scanning Confocal Microscopy ◽  
Apical Vesicles ◽  
Cytoplasmic Microtubules ◽  
Oriented Parallel

Urediospore germlings of Uromyces appendiculatus sense topographical signals inherent in host stomata over which they develop appressoria. The site of signal recognition by the fungus is invariably within the apical 6 μm of the germling. The organization of the germling apex before and during initiation of appressorium formation was examined by both electron microscopy and laser scanning confocal microscopy. Most notable were changes in the distribution and organization of the microtubules and apical vesicles. Microtubules were oriented parallel to the inductive ridge, along which expansion of the germling apex occurred. In germlings that contacted artificial inductive topographies in vitro, the distribution of apical vesicles in germlings did not change within the first 3–4 min. After 6–8 min of contact, most apical vesicles became repositioned near the ridge. The apical cluster in developing appressoria was closely associated with that population of cytoplasmic microtubules that developed parallel to the inductive ridge. Coincident with growth of the germling apex over an inductive ridge was a physical deformation of the cell by the ridge, indenting both the cell wall and plasma membrane. Key words: bean rust, cytoskeleton, infection structure, Spitzenkörper, urediospore.

scholarly journals Studying the Neurovascular Unit: An Improved Blood–Brain Barrier Model

2009 ◽  
Vol 29 (12) ◽  
pp. 1879-1884 ◽  
Author(s):  
Christoph M Zehendner ◽  
Heiko J Luhmann ◽  
Christoph RW Kuhlmann
Keyword(s):  
Blood Brain Barrier ◽  
Laser Scanning ◽  
Neurovascular Unit ◽  
Cell Types ◽  
Laser Scanning Confocal Microscopy ◽  
Brain Barrier ◽  
Scanning Confocal Microscopy ◽  
Blood Brain

The blood–brain barrier (BBB) closely interacts with the neuronal parenchyma in vivo. To replicate this interdependence in vitro, we established a murine coculture model composed of brain endothelial cell (BEC) monolayers with cortical organotypic slice cultures. The morphology of cell types, expression of tight junctions, formation of reactive oxygen species, caspase-3 activity in BECs, and alterations of electrical resistance under physiologic and pathophysiological conditions were investigated. This new BBB model allows the application of techniques such as laser scanning confocal microscopy, immunohistochemistry, fluorescent live cell imaging, and electrical cell substrate impedance sensing in real time for studying the dynamics of BBB function under defined conditions.

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