3D laser confocal microscope Keyence VK-X250 was used to measure the surface roughness of as-fabricated specimens. The imaging process takes surface height measurements using the point illumination method, where a laser beam is scanned across the sample in a raster pattern at incremental vertical displacements. The vertical resolution is dependent on the aperture of the objective lens. The measured height data was then analyzed using Keyence Multi-file Analyzer software. Image processing procedures were applied to account for the curved surface of the cylindrical samples, and surface roughness parameters are calculated. The surface roughness measurements were performed using an objective lens of 20 × magnification with a z-pitch of 0.20 µm. Approximately 4.7 mm × 1.4 mm of surface area was scanned within the central gage section of each specimen. The nominal scan step size was set to 1.4 µm which corresponds to 3345 × 1024 pixels within the surface map. Efforts were made to scan approximately similar regions across each specimen using print marks on specimens, as shown in Fig. 2 . Each specimen was aligned under the confocal microscope using print marks as guidance prior to scanning the central gage section.
Multifileanalyzer software
Manufactured by Keyence
Sourced in Japan
The MultiFileAnalyzer software is a data analysis tool developed by Keyence. It allows users to analyze and compare multiple data files simultaneously. The software provides functions for data visualization, processing, and statistical analysis.
Automatically generated - may contain errors
Lab products found in correlation
Vk x1000, by Keyence (4 mentions)
Vk x260k, by Keyence (2 mentions)
Vk x250, by Keyence (1 mentions)
Vk x1000 1050, by Keyence (1 mentions)
Teneo scanning electron microscope, by Thermo Fisher Scientific (1 mentions)
Vk x series, by Keyence (1 mentions)
Vk x200 series, by Keyence (1 mentions)
Vk x100, by Keyence (1 mentions)
12 protocols using multifileanalyzer software
1
Surface Roughness Analysis of Fabricated Specimens
2
Surface Roughness Evaluation of Materials
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A Keyence VK-X1000/1050 (Keyence Deutschland GmbH, Neu-Isenburg, Germany) confocal laser scanning microscopy (CLSM) with a Nikon CF IC EPI Plan 50X (NA: 0.5, NIKON, Osaka, Japan) objective was used for surface measurement. Sa as the amount of the height difference of each point compared to the arithmetic mean of the surface was evaluated with Multi File Analyzer software (2.1.3.89, Keyence Deutschland GmbH, Neu-Isenburg, Germany) according to ISO 25178-2:2012. Therefore, five different areas per samples were characterized five times each with an area of 100 µm × 100 µm each and filtered to receive the surface for roughness evaluation (S-Filter: 0.5 µm; F-Filter: 0.1 mm; Filter type: spline).
As values identified for Sa were not normally distributed across the samples of all materials analyzed in this study (Shapiro-Wilk test [30 (link)]: p < 0.05), medians and 25/75 percentiles were calculated, and statistical analyses were performed using the Kruskal–Wallis test [31 (link)] and post hoc analyses using the Dunn [32 (link)] post hoc test with Bonferroni [33 ] correction. To determine the strength of the effects, Cohen’s r [34 ,35 ] was calculated using the psychometrica platform. The general level of significance (α) was set to 0.05, and for interpretation, the Bonferroni-adjusted p-values were used.
As values identified for Sa were not normally distributed across the samples of all materials analyzed in this study (Shapiro-Wilk test [30 (link)]: p < 0.05), medians and 25/75 percentiles were calculated, and statistical analyses were performed using the Kruskal–Wallis test [31 (link)] and post hoc analyses using the Dunn [32 (link)] post hoc test with Bonferroni [33 ] correction. To determine the strength of the effects, Cohen’s r [34 ,35 ] was calculated using the psychometrica platform. The general level of significance (α) was set to 0.05, and for interpretation, the Bonferroni-adjusted p-values were used.
3
Detailed Plant Morphology Evaluation
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Observations were made four weeks after germination (10–12 replicates plants per genotype). Plant heights were measured manually from the soil level to the apical meristem, and the number of leaves per plant was counted. Compactness was calculated by dividing the number of leaves by the plant height. Leaf thickness was measured using a Mitutoyo 500 series digital caliper (Mitutoyo America Corporation, USA) by measuring thickness at five different points on the fifth leaf below the apical meristem and averaging these values per plant. Images of stomata were captured on both the abaxial and adaxial surfaces of the fifth leaf of four-week old plants under 20× magnification using a 3D laser scanning confocal microscope (VK-X260K, Keyence Corporation, USA). The imaging Z-range was adjusted for each sample in order to capture all the stomata in the field of view (712×534 µm2) that was being measured. Stomata were manually marked in the digital images, and then counted using MultiFileAnalyzer software (Keyence Corporation, USA). For each biological replicate, three separate images were captured from each leaf surface (abaxial and adaxial), and the numbers of stomata in these subsamples were averaged before analysis.
4
Imaging and Characterizing Surface Coatings
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Scanning electron microscopy (SEM) images were taken of the coated coverslips. Coatings were fixed in 3.7% glutaraldehyde and dehydrated using a series of increasing alcohol concentrations. Slides were then put in hexamethyldisilane and sputter coated in gold and silver. Images were taken using a Teneo scanning electron microscope (FEI, US).
Surface measurements to estimate film thickness were performed using a confocal laser scanning microscopy (VK-X series Keyence, Japan). Measurement mode was set to surface profile with a 20X lens magnification and measurement size set to super fine (2048 × 1536). Analysis of images obtained were carried out using Keyence MultiFileAnalyzer software.
Surface measurements to estimate film thickness were performed using a confocal laser scanning microscopy (VK-X series Keyence, Japan). Measurement mode was set to surface profile with a 20X lens magnification and measurement size set to super fine (2048 × 1536). Analysis of images obtained were carried out using Keyence MultiFileAnalyzer software.
5
Quantitative 3D Laser Scanning Analysis
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A 3D laser scanning confocal microscope VK-X1000 (Keyence, Japan) is used to image the prints and the Multifile analyzer software (Keyence, Japan) is used to analyze the images and measure the sizes of the patterns and structures obtained during the screen assembly and printing process. Three prints of each size and shape are taken, for each print six images are taken at 5X magnification to measure the horizontal dimensions of the prints and three images are taken at 20X magnification to measure the height of the prints. The diameters of the dots and the thickness of the lines and concentric circles of the printed structures are then measured using the plane measurement functions in the Multifile analyzer software and the vertical profile of the features are measured by using the profile function, and then the average height is calculated using the area under the profile curve and the horizontal thickness of the feature.
6
Microscopic Analysis of Coagulated Dairy Blends
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An aliquot of each coagulated CM:PP blend was placed on a 3-mm microscope slide and gently dried at 37°C in an oven. Afterward, it was analyzed with a 3-dimensional laser microscope (VK-X200 series, Keyence) under different magnifications (10×, 20×, 50×, and 150× lenses). Four images were analyzed for each sample, and the most representative was chosen and depicted for analysis. Five areas of each image were analyzed to determine surface roughness (Sa; as the arithmetical mean height in μm) using Multi File Analyzer software (version 1.3.1.120; Keyence). The averages of each CM:PP blend Sa values were compared with Sa values of the positive control (coagulated skim milk).
7
Fabricating Grooved Polystyrene Substrates
8
Microscopic Analysis of Hair Surface Texture
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A
confocal Keyence VK-X150 microscope (equipped with long-distance
100× Nikon objective lens) was used for the morphological investigations
of hair surface texture. Hair segments were attached to glass slides,
similarly to AFM (as described above). Laser and optical images were
processed using VK Analyzer software (v.3.8.0.0) to remove tilt and
reduce noise. Morphological analyses were conducted on hair segments
treated by keratin and halloysite/keratin. For comparison, confocal
images of uncoated hair samples were collected. Surface texture analysis
was performed using a Keyence MultiFileAnalyzer software (v.1.3.0.116).
confocal Keyence VK-X150 microscope (equipped with long-distance
100× Nikon objective lens) was used for the morphological investigations
of hair surface texture. Hair segments were attached to glass slides,
similarly to AFM (as described above). Laser and optical images were
processed using VK Analyzer software (v.3.8.0.0) to remove tilt and
reduce noise. Morphological analyses were conducted on hair segments
treated by keratin and halloysite/keratin. For comparison, confocal
images of uncoated hair samples were collected. Surface texture analysis
was performed using a Keyence MultiFileAnalyzer software (v.1.3.0.116).
9
Surface Characterization of PVP-Coated Polymer
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A 3D Keyence Laser Microscope (LSCM, VK-X260K, Keyence, USA) was used to evaluate the surface morphology and topography of the guide. 3D measurement data were analyzed with Keyence’s Multi-File Analyzer software. Samples were examined in the following order: polymer laminate before saturation with PVP, polymer laminate after saturation with PVP. Both sets of samples were examined using 20X lens and 100X lens.
10
Foam Geometry and Melting Layer Analysis
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To investigate the influence of the TAFP parameters on the change of the foam geometry, the initial height hi and the height hc of the compressed foam specimens (Figure 6 ) are measured with a Holex ABS calliper (Hoffmann SE, Munich, Germany) with a resolution of 0.01 mm. Since the test specimens are measured just before and after the compression test, only the plastic foam deformation is recorded, and therefore the elastic foam deformation is not considered in this study. In addition, three lateral optical microscope images (Keyence VK-X1000 with 2.5× magnification) of each of the compressed foam specimens at regular distances across the specimen width are taken to determine the thickness of the forming melting layer tl. First, the transition between the melting layer and the undeformed foam structure is identified (dotted line in Figure 6 ). Subsequently, the distance between the identified transition within the foam test specimen and the foam test specimen surface (corresponding to the thickness of the melting layer tl) is determined by using parallel compensation lines within the Keyence Multi-File-Analyzer® software. Finally, an average melting layer thickness tm is calculated for each parameter set. For error estimation of the determined foam deformation and melting layer thickness, the standard deviation is used in each case.
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