An aliquot of 40 μL running buffer (10 mM Tris-HCl, 0.05% v/v Tween 20, pH 7.0) was mixed with 0.1μL of the AuNPs-biotinyl-αIL6ab conjugates, and the solution mixture was then applied to the streptavidin coated LFIDs and GFC-LFIDs and allowed to run for 10 min. The assay was completed by further addition of 20 μL of the running buffer. For quantitative evaluation, images of the GFC-LFIDs were recorded using a flatbed scanner (Epson Perfection V370 Photo). The color intensities of the test zones were analyzed with ImageJ software (Scion Corp., USA) and quantified as a function of pixel intensity.
Perfection v370 photo
Manufactured by Epson
The Epson Perfection V370 Photo is a flatbed scanner designed for scanning photographs, documents, and other flat items. It features a 4800 dpi optical resolution and can scan items up to 8.5 x 11.7 inches in size.
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10 protocols using perfection v370 photo
1
Quantitative Evaluation of AuNP-Biotin-αIL6 Assay
2
Digit Ratio Measurements Using Scanner
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We scanned the participants’ right and left hands using an Epson Perfection V370 Photo scanner in gray level with 300 DPI resolution. The participants were instructed to remove all jewelry from their hands, slightly spread the fingers, and have contact to the scanner with every finger segment. We used the GNU Image Manipulation Program (GIMP; www.gimp.org ) to quantify the length of the second (2D) and fourths (4D) digits, i.e. distance from the middle of the basal crease to the tip of the fingers. Three independent raters (RBJ, BA, AS) measured each finger three times (nine times in total) and were uninformed about sex, age, and brain volumes. We defined M2D:4D as our primary predictor. R2D:4D and L2D:4D were also tested as further predictors. The inter-rater reliabilities (two-way random inter-rater correlation coefficient; absolute agreement) were very high: M2D:4D: n = 61, 0.969; R2D:4D: n = 61, 0.956; L2D:4D: n = 61, 0.963.
3
Quantifying A549 Cell Colony Formation
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At 24 h post-transfection, A549 cells were harvested and manually counted under an optical microscope. The cells (6x102/well) were subsequently seeded into a 6-well plate, and cultured in RPMI 1640 medium (with 10% FBS) in a humidified incubator at 37˚C for 10-14 days. The culture media were replaced every 2 days until the end of the experiment. Finally, the cells were fixed with 4% paraformaldehyde for 30 min at room temperature (RT), and then stained with 0.1% crystal violet at RT for 1 h. The plates were dried and scanned using a Epson Perfection V370 Photo scanner. Cell colonies (>50 cells) were counted manually.
4
Tomato Cultivation and NMR Spectroscopy
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NMR spectra were recorded on an AVANCE III spectrometer (400 MHz). The solvent residue was referred to by the chemical shifts (in ppm). Coupling constants (J) were recorded in Hertz. LC-MS/MS data were read on an AB Sciex Triple TOF 5600+ instrument (AB Sciex Pte. Ltd., Framingham, MA, USA). Tomatoes were grown in an artificial climate chamber. Plant photos were scanned by an Epson Perfection V370 Photo. Chemicals were bought from Energy Chemical Co. Ltd. (Shanghai, China) without further purification before use.
5
Lateral Flow Immunoassay for IL-6 Quantification
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An aliquot of 40 μL sample solution with various interleukin-6 (IL-6) concentrations ranging from 0.1 to 10 ng/mL was mixed with 1 μL of the AuNPs-biotinyl-αIL6ab conjugates, and the solution mixture was then applied to the primary anti-interleukin-6 antibodies coated LFIDs and GFC-LFIDs and allowed to run for 10 min. The assay was completed by further addition of 20 μL running buffer (10 mM Tris-HCl, 0.05% v/v Tween 20, pH 7.0). For quantitative evaluation, images of the LFIDs and GFC-LFIDs were recorded using a flatbed scanner (Epson Perfection V370 Photo). The color intensities of the test zones were analyzed with ImageJ software (Scion Corp., USA) and quantified as a function of pixel intensity. The images were converted to grey levels, and a region of interest along the strip axis intercepting the test line was established. The amplitude of the intensity profile measured with respect to the base line (membrane apart from the test line) was used to quantify the response intensity.
6
Automated Colony Tracking and Quantification
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Overnight cultures were diluted in LB or M9 media 1:1000. Cells were grown up to an OD of 1 before plating at appropriate dilutions on solid agar medium. Standard PC scanners (i.e., Epson Perfection V370 Photo) were used to acquire images at 800 dpi resolution of Petri dishes every 10 min. Acquired images were analyzed with an in-house image analysis software implemented in ImageJ IJ1 Macro and Python. In order to improve the accuracy in detection of bacterial colonies the software corrects for changes in light across plates (i.e., Otsu thresholding over each plate), performs colony segmentation (i.e., watershed algorithm over the standard variation of the experiment’s Z-stack projection of all images) and tracking. Finally, outliers/artifacts are filtered out. The final data returns for each colony x and y coordinate on the plate, size at each time point, and RGB intensity. Full details and computer codes are available on GitHub https://github.com/Dfernand1795/PetriScanner2 .
7
Oil Red O Staining of Adipocytes
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Differentiated adipocytes were washed twice with PBS, fixed with 4% paraformaldehyde at room temperature for 15 min and washed twice again with PBS. Then, cells were stained with 5 mg ml−1 oil red O in isopropanol (O0625, Sigma-Aldrich) at room temperature for 2 h. After that, cells were washed three times with tap water and left to dry at room temperature. For visualizing the scanner Epson Perfection V370 Photo was used.
8
Colony-forming Assay with Wright-Giemsa Staining
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Colony-forming assays were performed using Wright-Giemsa stain (Baso, CHN) following manufacturer's instructions. Briefly, cells were seeded into 24-well microplates and sorafenib was added after 24h of incubation. The cells were cultured for another 48h, and 300μl Wright-Giemsa stain was added to each well. The microplates were incubated at 37°C for 1 min. Then, we rinsed it with double-distilled water gently for three times, dried and examined the finished slide under a microscope. Finally, the 24-well microplates were photographed by using scanner (Epson Perfection V370 Photo).
9
Root Imaging and Morphometry Analysis
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Roots were thoroughly washed out and stained with an abundant volume of 0.01% neutral red (Sigma Chemical Co.) for 24 hours to increase contrast for further analysis (Schumacher et al., 1983) . The stained roots were placed in a transparent tray with a thin layer of water and scanned using a commercial scanner (Epson Perfection V370 Photo) at a resolution of 600 pixels per mm.
Root images were analyzed using WinRHIZO (Regent Instruments Inc., Québec City, QC, Canada) as described by Himmelbauer et al. (2004) and total root length, surface, volume, average diameter, number of forks and root length per diameter class were recorded.
Root images were analyzed using WinRHIZO (Regent Instruments Inc., Québec City, QC, Canada) as described by Himmelbauer et al. (2004) and total root length, surface, volume, average diameter, number of forks and root length per diameter class were recorded.
10
Predicting Total Leaf Area in Trees
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Assessment of total leaf area (TLA) of mature trees is labor intensive and a reliable predictor of TLA facilitates evaluation of the relationships to growth and physiological traits.
To identify a reliable predictor for TLA we conducted a pilot study where seven trees were harvested in Pustnäs (Sweden) in July 2013. Altogether we harvested seven plants consisting of 84 shoots in total with a height above 1 m. The following parameters were tested: leaf areas within different shoot sections (within 20, 30, 40 and 50 cm) along the main shoot, main shoot height, main shoot diameter, main shoot crown length, number of shoots, total crown height, total height, total leaf biomass, total diameter at 1.20 m height (Table 2). Sapwood area (SA) was calculated assuming circular shoot cross-sections and that sapwood extended to the center of the shoot (a reasonable assumption for these young shoots). Specific leaf area (m 2 g -1 ) was analysed by assessing individual leaf areas with a flat-bed scanner (Epson Perfection V370 Photo) and the open-source software ImageJ (Yang Yu et al. 2011) (link), and the subsequent determination of leaf dry weights after drying the leaves for 48 h at 70 °C. In September 2013, additionally 14 trees were sampled in Italy for determination of TLA and stem diameter.
To identify a reliable predictor for TLA we conducted a pilot study where seven trees were harvested in Pustnäs (Sweden) in July 2013. Altogether we harvested seven plants consisting of 84 shoots in total with a height above 1 m. The following parameters were tested: leaf areas within different shoot sections (within 20, 30, 40 and 50 cm) along the main shoot, main shoot height, main shoot diameter, main shoot crown length, number of shoots, total crown height, total height, total leaf biomass, total diameter at 1.20 m height (Table 2). Sapwood area (SA) was calculated assuming circular shoot cross-sections and that sapwood extended to the center of the shoot (a reasonable assumption for these young shoots). Specific leaf area (m 2 g -1 ) was analysed by assessing individual leaf areas with a flat-bed scanner (Epson Perfection V370 Photo) and the open-source software ImageJ (Yang Yu et al. 2011) (link), and the subsequent determination of leaf dry weights after drying the leaves for 48 h at 70 °C. In September 2013, additionally 14 trees were sampled in Italy for determination of TLA and stem diameter.
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