No abstract available.
Microscopy, Confocal

Confocal laser endomicroscopy technology can obtain cell-level images in real time and in situ, which can assist doctors in real-time intraoperative diagnosis, but its non-invasiveness makes it difficult to relocate the optical biopsy site. The confocal probe localization algorithm can automatically calculate the coordinates of the probe tip, that is, the coordinates of the optical biopsy site. In this paper, a confocal probe localization algorithm based on region growing and endoscope size prior was proposed. The algorithm detected the probe region by region growing on the probe edge image, then searched for tip points based on a given probe axis, and iteratively optimized it. Finally, based on the single-degree-of-freedom motion characteristics of the probe, the three-dimensional coordinates of the tip of the probe were calculated by using the prior information of the size of the endoscope, which solved the scale uncertainty problem of the monocular camera. The confocal probe localization algorithm was tested on the dataset collected in this paper. The results showed that our algorithm no longer relied on the color information of the probe, avoided the influence of uneven illumination on the gray value of the probe pixels, and had a more robust location accuracy and running speed. Within the length of the probe extending out of the endoscope from 0 to 5 cm, the pixel error could be as low as 11.76 pixels, and the average relative position error could be as low as 1.66 mm, which can achieve the real-time and accurate localization of the confocal probe.
Endoscopes ; Algorithms ; Microscopy, Confocal/methods*

PURPOSE: In this study we analyzed and objectified the characteristics of the Avellino corneal dystrophy patients considering disease severity using in vivo confocal microscopy (IVCM). METHODS: Each corneal layer of 36 eyes in 18 patients with Avellino corneal dystrophy was examined using IVCM (ConfoScan 4.0, NIDEK, Co. Ltd., Albignasego, Italy). Patients were classified into 3 groups based on disease severity (mild, moderate, or severe). RESULTS: In the mild group, hyper-reflective granular deposits without dark shadows were observed in the anterior stroma. As the disease progressed, corneal deposits were also found at the posterior stroma and epithelium, and clusters of hyper-reflective corneal deposits resembling stromal opacity were noted. The range of corneal deposits measured using Z-scan optical pachymeter was 111.14 +/- 30.95 um in the mild group, 157.47 +/- 25.00 um in the moderate group, and 193.42 +/- 52.23 um in the severe group (p < 0.05). CONCLUSIONS: The origin of the corneal deposits in Avellino corneal dystrophy may be related to corneal stromal cells and distributed from the corneal epithelial layer to the stromal layer. IVCM might be useful for the standardization of disease severity by digitalizing the range of deposits.
Epithelium ; Humans ; Microscopy, Confocal ; Stromal Cells

OBJECTIVES: The purpose of this study was to compare the cutting efficiency of a newly developed microprojection tip and a diamond-coated tip under two different engine powers. MATERIALS AND METHODS: The apical 3-mm of each root was resected, and root-end preparation was performed with upward and downward pressure using one of the ultrasonic tips, KIS-1D (Obtura Spartan) or JT-5B (B&L Biotech Ltd.). The ultrasonic engine was set to power-1 or -4. Forty teeth were randomly divided into four groups: K1 (KIS-1D / Power-1), J1 (JT-5B / Power-1), K4 (KIS-1D / Power-4), and J4 (JT-5B / Power-4). The total time required for root-end preparation was recorded. All teeth were resected and the apical parts were evaluated for the number and length of cracks using a confocal scanning micrscope. The size of the root-end cavity and the width of the remaining dentin were recorded. The data were statistically analyzed using two-way analysis of variance and a Mann-Whitney test. RESULTS: There was no significant difference in the time required between the instrument groups, but the power-4 groups showed reduced preparation time for both instrument groups (p < 0.05). The K4 and J4 groups with a power-4 showed a significantly higher crack formation and a longer crack irrespective of the instruments. There was no significant difference in the remaining dentin thickness or any of the parameters after preparation. CONCLUSIONS: Ultrasonic tips with microprojections would be an option to substitute for the conventional ultrasonic tips with a diamond coating with the same clinical efficiency.
Dentin ; Diamond ; Microscopy, Confocal* ; Tooth ; Ultrasonics*

OBJECTIVEMicrostructural changes in dentin carious lesions were investigated using a 3D laser scanning microscope, which has a morphological theoretical foundation in the further study of clinical caries disease prevention and treatments.

METHODSSix fresh extracted caries molars were prepared into cross-section specimens. The sections were examined by 3D and laser measuring morphology.

RESULTSZones were identified in the lesions on the basis of their optical appearance. Two zones were identified in the lesions on the basis of their laser appearance. The microstructure showed that the tubular was partly closed in transparent dentin; peritubular and intertubular dentin were reduced in the zone of demineralization; peritubular and intertubular dentin were damaged and fused; a beaded sample and oval lesions formed in the zone of bacterial invasion; and abnormal dentin structure was present in the zone of destruction on the basis of their laser appearance. Four zones were iden-tified in the lesions according to their colors, as determined from their 3D appearance.

CONCLUSIONS3D laser scanning micros-cope may be a powerful, accessible, and non-destructive technique, as it identified the lesion and tubular zones, as well as peritubular and intertubular dentin in the four zones' lesions. The microstructure of dentin caries lesions may have significant merit in the evaluation of clinical prevention and treatment.

Although traditional tooth preparation techniques (e.g., depth-groove-guided and index-guided techniques) are designed to improve preparation precision, the results are unsatisfactory because of the lack of proper estimating tools. This study proposed a novel technique, in which relevant details for preparation of drilling holes are provided and corresponding depth is estimated using a quantitive bur under a microscope. This technique offers a viable option for precise tooth preparation.
Humans ; Microscopy, Confocal ; Tooth Preparation ; methods

The purpose of this study was to evaluate the effects of a sealant resin on enamel demineralization in orthodontic bracket bonding. The forty eight extracted sound bovine teeth were subdivided into four groups and treated with Phase II(R) (Reliance, Itasca, Ill) on the surface. Group 1 was not treated. Group 2 was acid etched with 37% phosphoric acid for 30 seconds. Group 3 was applied with sealant after acid etching. Group 4 was applied with resin paste after acid etching and sealant application. Each group was demineralized in artificial caries solution. Demineralized enamel depth was measured by confocal laser scanning microscopy. The results were as follows: the mean demineralized enamel depth was 47.4 micrometer (Group 1), 61.8 micrometer (Group 2), 13.9 micrometer (Group 3), 8.2 micrometer (Group 4); the demineralized enamel depth was increased in Group 2 than in Group 1 (p < 0.05); the demineralized enamel depth was reduced in Group 3 than in Group1 and Group 2 with statistically significant differences (p < 0.05); and demineralization in Group 4 was very little. The results of the present study indicate that sealant application is useful for reducing enamel demineralization in orthodontic bracket bonding.
Dental Enamel* ; Microscopy, Confocal ; Orthodontic Brackets* ; Tooth

PURPOSE: To compare the results of specular microscopy with those of confocal microscopy for evaluation of corneal endothelium. METHODS: We evaluated corneal endothelium of 103 eyes using specular microscopy and confocal microscopy. Endothelial cell density, pleomorphism, and polymegathism were measured using a ConfoScan 4 confocal microscope (Nidek Technologies, Inc, Greensborom, NC) in automatic mode before and after manual correction. Also, endothelial cell density, the coefficient of variation, and hexagonality were evaluated using a Konan Noncon Robo-8400 noncontact specular microscope (Konan medical, Inc., Hyogo, Japan). The differences in results obtained from these various methods were compared: polymegathism was compared with the coefficient of variation, and pleomorphism was compared with the inversion of hexagonality. RESULTS: Endothelial cell density as measured by specular microscopy, the automatic count of confocal microscopy, and the manual correction for confocal microscopy were 2797.6+/-354.14 cell/mm2, 2973.1+/-284.24 cell/mm2, and 2861.9+/-335.58 cell/mm2, respectively. Results of each test was not significantly different (p=0.241). The inversion of hexagonality, pleomorphism of automatically counted confocal microscopy, and the pleomorphism of manually corrected confocal microscopy were 56.14%, 54.77%, and 55.24%, respectively. Results of each test were not significantly different (p=0.147).The coefficient of variation of specular microscopy, the polymegathism of automatic counted confocal microscopy, and the polymegathism of manually corrected confocal microscopy were 33.71%, 39.68%, and 38.75%, respectively. Results of each test were significantly different (p=0.005). CONCLUSIONS: Endothelial cell density and polymegathism as measured by confocal microscopy were not different from specular microscopy results in normal corneas, but these results were different for polymegathism in normal corneas. Therefore, manual correction for endothelial cell evaluation of a disordered cornea should be performed during clinical evaluation.
Cornea ; Endothelial Cells ; Endothelium, Corneal ; Eye ; Microscopy ; Microscopy, Confocal

Dendritic cells (DCs) are professional antigen presenting cells of the immune system. They can be generated in vitro from peripheral blood monocytes supplemented with GM-CSF, IL-4 and TNF alpha. During induction, DCs will increase in size and acquire multiple cytoplasmic projections when compared to their precursor cells such as monocytes or haematopoietic stem cells which are usually round or spherical. Morphology of DCs can be visualized by conventional light microscopy after staining or phase-contrast inverted microscopy or confocal laser scanning microscopy. In this report, we described the morphological appearances of DCs captured using the above-mentioned techniques. We found that confocal laser scanning microscopy yielded DCs images with greater details but the operating cost for such a technique is high. On the other hand, the images obtained through light microscopy after appropriate staining or phase contrast microscopy were acceptable for identification purpose. Besides, these equipments are readily available in most laboratories and the cost of operation is affordable. Nevertheless, morphological identification is just one of the methods to characterise DCs. Other methods such as phenotypic expression markers and mixed leukocyte reactions are additional tools used in the characterisation of DCs.
Dendritic Cells/*cytology ; Microscopy, Confocal ; Microscopy, Phase-Contrast

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