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676 protocols using illustrator cs6

1

Skin Graft Contraction and Appearance

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For > 6 months after the procedure, the recipient site was observed for graft take, presence of secondary contraction, and color and texture match. Secondary contraction in particular was checked by tracing and scanning the graft skin immediately after the procedure and at 6 months (or more) after the operation and outlining the data on Adobe Illustrator CS6. A plug-in computer software “Hakariya” (Comnet Co., Ltd., Kobe, Japan), which can be used for automatically calculating square measure, circumference and length, and settings of scales on Adobe Illustrator CS6 was downloaded. The square measures of the graft skin were calculated according to the set scale, thus contraction rate was confirmed (Fig. 1). Contraction percentages were defined as follows: <5% : “no contraction,” 5%–30% : “mild contraction,” and >30% : “contraction.”
Regarding color and texture match of the recipient site, results were defined as follows: good: mild pigmentation/both patient and physician are satisfied; fair: mild pigmentation/either patient or physician is dissatisfied; and poor: severe pigmentation.
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2

Optimized Visualization of Protein Complex Profiles

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Figures were prepared using the programs Affinity Designer 2 (version 2.1.1) and Adobe Illustrator CS6 (version 16.0.4). The 2D gels of Figures 1A and 4G were mirrored for the consistency of presentation. The MitCOM profiles were derived from the dataset of Schulte et al.33 (link) and https://www.complexomics.org/datasets/mitcom (Gaussian smoothing 1 and quality control of profiles/peptides) and graphs were prepared using the programs GraphPad Prism 9 (version 9.0.0) and Adobe Illustrator CS6 (version 16.0.4).
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3

Cytotoxic effects of novel compounds

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All values were expressed as mean ± SD. Data were analyzed by one-way variance (ANOVA) followed by Tukey’s multiple comparison with SPSS 25 software (IBM SPSS Statistics 25.0, Armonk, NY, USA). The p < 0.05, p < 0.01 and p < 0.001 were set at the threshold for statistical significance, high statistical significance, and very high statistical significance, respectively. Graphs were constructed using GraphPad Prism 8 (GraphPad Prism 8.0, San Diego, CA, USA) and Adobe illustrator CS6 software (Adobe illustrator CS6, San Jose, CA, USA).
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4

Kaplan-Meier Survival Analysis of NSCLC and ADC

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Kaplan-Meier survival curves with hazard ratio and log-rank p value were calculated and plotted with the analysis tool, which can be accessed online at http://kmplot.com/analysis/ [35 (link)]. The background database, downloaded from GEO (Affymetrix microarrays only), EGA, and TCGA, offers gene expression data, relapse free survival (RFS), and overall survival (OS) rates. The software is capable to assess the effect of 54,675 genes on survival using 10,188 cancer samples, among which includes 2437 cases of lung cancer. We analyzed the survival outcomes of NSCLC and ADC in different expression levels of DACH1 and CXCL8. The Kaplan-Meier survival curves downloaded from the website were resized in Adobe Illustrator CS6. The blend curves were obtained from IBM SPSS Statistics 19.0 using GSE31210 and resized in Adobe Illustrator CS6.
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5

Multimodal Imaging of Mouse Brain

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Immunohistochemistry was performed according to methods described previously (17 (link)). For EdU detection, the Click-iT EdU Alexa Fluor 555 imaging kit (Thermo Fisher Scientific) was used, and slices were treated according to the manufacturer’s instructions. Immunohistochemical stainings were imaged with a confocal microscope [Leica SP5 or SP8; 20× numerical aperture (NA), 0.7; 40× NA, 1.25; and 63× NA, 1.4] or stereomicroscope (Zeiss Axio Zoom.V16; 1× NA, 0.25). For stains that exhibited salt-and-pepper noise, a median filter of 5 × 5 × 5 was applied to eliminate noise. Images were analyzed using the image processing software Imaris 9.2.0. (Bitplane) and Adobe Illustrator CS6.
For histochemistry, mouse brains were collected and embedded in paraffin. Two-micrometer-thick tissue sections were stained with PAS, hematoxylin and eosin, or the Bielschowsky silver stain using standard protocols. For Alizarin red staining, sections were deparaffinized and rehydrated, as well as incubated for 1 hour in 1% Alizarin red solution (pH 9.0) followed by 1 hour in 1% Alizarin red solution (pH 6.4) at room temperature. Stained paraffin sections were scanned with NanoZoomer HT (Hamamatsu Photonics), equipped with a 20× objective (UPlanSApo; NA, 0.75; Olympus). Images were analyzed using Digital Image Hub software (SlidePath) and Adobe Illustrator CS6.
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6

Vector Image and Data Visualization Protocol

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All vector maps (Figures 1 and 2, and Supplementary Figure S1) were created using Vector NTI Advance 11.5.0 (Thermo Fisher Scientific, Inc.), exported as .wmf files and annotated using Adobe Illustrator CS6 (Adobe). Digital agarose gel pictures (.tif format) were imported into Adobe Photoshop CS6 (Adobe), exported in Adobe Illustrator format and subsequently annotated using Adobe Illustrator CS6. Figure 3 was generated in Prism 6 (GraphPad) based on data from Table 1, and annotated in Adobe Illustrator. Figure 4 was generated entirely in Adobe Illustrator.
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7

Confocal and Electron Microscopy Imaging

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Samples for confocal laser scanning microscopy (antibody staining and in situ hybridization) were mounted in Murray’s clear and scanned in either Leica SP5 or Olympus FV3000 CLSM. Z-stacks of confocal scans were projected into 2D images in IMARIS 9.1.2. TEM microphotographs were obtained with Gatan ES500W camera mounted on transmission electron microscope Jeol JEM-1011. Both CLSM images and TEM micrographs were assembled in Adobe Illustrator CS6 into final figures. All the schematic drawings were done with Adobe Illustrator CS6.
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8

Detailed Visualization and Statistical Analysis Methods

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Illustrations were prepared using PowerPoint (Microsoft) and Illustrator CS6 (Adobe). Bar graphs, dose-response graphs and circle graphs were prepared using Prism 7 (GraphPad Software) and statistical analysis was performed using the same software. For comparisons between two datasets, significance was determined by unpaired t test; for comparisons between more than two datasets, significance was determined by one-way ANOVA. Significance is indicated as **** (p<0.0001), ** (p<0.01), * (p<0.05) or ns (not significant). Pictures of gels and immunoblots were only adjusted for contrast and brightness using Photoshop CS6 (Adobe) and were arranged in Illustrator CS6.
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9

Motoneuron Labeling and Visualization

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Sections for LM were examined with a Leica DMR microscope equipped with a digital camera (Leica DFC-450). Photopanels were constructed with Adobe Photoshop and Illustrator CS6. The 3D-plots of the spinal cords of Figures 3D1–3 were made using a motorized Olympus BH microscope equipped with a Lucivid miniature monitor and NeurolucidaTM software (Microbrightfield). Cells were determined to be motoneurons if they were located within the lateral motoneurons column, had a diameter of at least 25 μm. Cells were only plotted if they contained a nucleus. RABV deposits reflecting lytic motoneurons were plotted if the size of the deposit was at least 40 μm. The Neurolucida set-up was also used to count and measure contours of labeled neurons. Diagrams were constructed with ExcelTM (Microsoft Office 2010).
Fluorescent labeling was assessed using either a Leica DMRBE microscope and appropriate filters and photographed with a Hamamatsu camera (C4880) or with a Zeiss LSM700 confocal microscope and Zeiss 2009 software (ZenTM). Photopanels were constructed in Adobe Photoshop and Illustrator CS6.
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10

Image Processing and Visualization

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Z-stacks of confocal scans were projected into 2D images and 3D reconstructions in IMARIS 9.1.2. Both light micrographs and CLSM images were adjusted in Adobe Photoshop CS6 and assembled in Adobe Illustrator CS6. All the schematic drawings were done with Adobe Illustrator CS6.
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