Two previously calibrated examiners (A and B) assessed dye penetration in the gingival margin visually, under a stereomicroscope (Model SZX7, Olympus Corporation, Tokyo, Japan), with ×20 magnification, and evaluated digital images, by using the Image Tool Software 3.0® (ITS), which is of public domain. Each examiner performed two evaluations in two different occasions, independently, and under blind conditions. The interval between the two readings was 15 days. Subjectivity was controlled by means of the pre-test study previously described, where the examiners evaluated teeth fragments under stereomicroscope. Intra- and inter-examiner agreement was obtained according to the visual and digital methods, as follows.
Model szx7
Manufactured by Olympus
Sourced in Japan, United States
The Olympus Model SZX7 is a stereomicroscope designed for routine observation and analysis. It features a zoom magnification range and provides a stereoscopic, three-dimensional view of the specimen.
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7 protocols using model szx7
1
Evaluating Gingival Margin Dye Penetration
2
Transgenic Mouse Model of Spinocerebellar Ataxia
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The MJD transgenic mice used in this study were previously described [21 (link)]. CMVMJD135 mice express the human ATXN3 carrying 135 glutamines and develop a progressive neurological phenotype with onset at 6 weeks, which includes loss of strength, impairment of motor coordination, loss of balance and altered reflexes. At late stages they show an overall reduction in brain weight, with reduced volume and/or total cell number in pontine nuclei and deep cerebellar nuclei, and intranuclear ATXN3 inclusions in several disease-relevant regions of the brain and spinal cord. Transgenic mice and control non-transgenic littermate mice (n = 4–5 pools of two animals per genotype) with a mean age of 20 weeks were sacrificed by decapitation, and brain slices were obtained for the macrodissection of the deep cerebellar nuclei using a stereomicroscope (Model SZX7, Olympus America Inc., USA) and frozen at −80ºC. For immunofluorescence assays, transgenic and wild-type littermate mice (mean age: 24 weeks) were deeply anesthetized and transcardially perfused with sterile PBS followed by 4% paraformaldehyde (PFA) in PBS. Brains were post-fixed overnight in fixative solution and embedded in paraffin. Slides with 4-μm-thick paraffin sections were processed for immunostaining with anti-PKCδ, -ATXN3 and—PNKP antibodies.
3
Huntington's-like Mouse Model for Ataxia Studies
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CMVMJD135 mice, expressing human ataxin-3 carrying 135 glutamines, were used in this study [37 (link)]. These mice display a progressive motor phenotype starting at an age of 6 weeks, with extensive phenotypic overlap with the human disease; they also develop ATXN3-positive neuronal inclusions in different regions of the brain and spinal cord, as well as a cell number and/or volume decrease in key regions for the disease, such as the pontine nuclei and the dentate nuclei of the cerebellum. Transgenic mice and control non-transgenic littermate mice (n = 5 per genotype) with a mean age of 25 weeks were sacrificed by decapitation, and brain slices were obtained for the macrodissection of pontine nuclei, substantia nigra, deep cerebellar nuclei and hippocampi using a stereomicroscope (Model SZX7, Olympus America Inc., Center Valley, PA, USA). Nuclear extracts from these different brain regions were obtained as previously described [62 (link)].
4
Detailed Dissection and Morphology of G. koreana
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The specimens were transferred to glycerine for dissection, and then examined and dissected under a dissection microscope (Olympus, model SZX-7). Figures of dissected appendages were drawn under a light microscope with an attached drawing tube (Leica, model DM 2500). Figures of the whole body were drawn using a drawing tube attached to a stereomicroscope (Olympus, model SZX-12). The lengths of all appendages and the whole body were measured with a stage micrometre (Leica, model no. 11513106) and an ocular micrometre. The photograph of the whole body of G. koreana
Morphological terminology and the orientation of each appendage largely follows Bruce (2009) ; some morphological terms were taken from Cohen and Poore (1994) (link) to retain descriptive consistency for the cephalic appendages ofG. koreana
Morphological terminology and the orientation of each appendage largely follows Bruce (2009) ; some morphological terms were taken from Cohen and Poore (1994) (link) to retain descriptive consistency for the cephalic appendages of
5
Dissection and Microscopic Examination
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The specimens were dissected in 70 % ethanol and mounted in gum-chloral medium on glass slides under a stereomicroscope (Model SZX-7; Olympus, Tokyo, Japan). Specimens were examined using a Nikon Eclipse Ni light microscope (Nikon, Tokyo, Japan) and illustrated with the aid of a drawing tube. The body length from the tip of the rostrum to the base of the telson was measured along the dorsal curvature to the nearest 0.1 mm. The nomenclature of the setal patterns on the mandibular palp follows that of Stock (1974) .
6
Morphological Characterization of RCF
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Morphology of RCF was analyzed in a stereomicroscope system SZX7 model from Olympus (Tokyo, Japan) using an ocular magnifying glass of 50×. This was equipped with a KL1500-LCD light source. Fracture surfaces of the pieces after the impact tests were observed by FESEM in a CARL ZEISS Ultra-55 FESEM microscope from Oxford Instruments (Abingdon, UK). To provide conducting properties, the samples were previously covered with a with a 5–7 nm gold-palladium layer in vacuum conditions in a cathodic sputter-coater Emitech SC7620 from Quorum Technologies LTD (East Sussex, UK). The acceleration voltage during FESEM analysis was set to 2.0 kV.
7
Optical and Electron Microscopy of PLA and FFs
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Optical microscopy of the as-received FFs was performed using a stereomicroscope system SZX7 model from Olympus (Tokyo, Japan) with an ocular magnifying glass of 12.5× and equipped with a KL1500-LCD light source. The morphology of the as-received FFs and the fracture surfaces of the injection-molded PLA and PLA/FF pieces obtained from the impact strength test were also observed by field emission scanning electron microscopy (FESEM) in a ZEISS ULTRA 55 FESEM microscope from Oxfrod Instruments (Abingdon, UK). The microscope worked at an acceleration voltage of 2 kV. Prior to analysis, the fracture surfaces were coated with a gold-palladium alloy in a Quorun Technologies Ltd. EMITECH mod. SC7620 sputter coater (East Sussex, UK).
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