KYSE70, KYSE450, or KYSE510 cells were suspended in complete growth media and then 0.3% agar with different doses of xanthohumol was added in a top layer over a base layer of 0.5% agar. The cultures were maintained at 37°C in a 5% CO2 incubator. Three weeks later, the colonies were counted and photographed under a microscope using the Image‐Pro Plus software (v.6.0) program (Media Cybernetics, Rockville, MD).
Image pro plus software v 6.0
Manufactured by Media Cybernetics
Sourced in United States
Image-Pro Plus software (v.6.0) is a comprehensive image analysis and processing software. It provides a suite of tools for capturing, processing, analyzing, and managing digital images. The software is designed to work with a variety of image formats and supports multiple image acquisition devices.
Automatically generated - may contain errors
Lab products found in correlation
Optima 5300 dv, by PerkinElmer (2 mentions)
Jem 2100, by JEOL (2 mentions)
Fv10 asw software v 4, by Olympus (2 mentions)
Fv1000 confocal microscope, by Olympus (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Dxm 1200 camera, by Nikon (1 mentions)
Calreticulin, by Cell Signaling Technology (1 mentions)
Emccd camera, by Olympus (1 mentions)
Bx53 microscope, by Olympus (1 mentions)
Ab18256, by Abcam (1 mentions)
Bs084, by Biosharp (1 mentions)
Bs114, by Biosharp (1 mentions)
Dylight 488 goat anti rabbit igg, by Abbkine (1 mentions)
A23220, by Abbkine (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Inverted light microscope, by Olympus (1 mentions)
Abs130868, by Absin (1 mentions)
Trans blot turbo mini pvdf transfer pack, by Bio-Rad (1 mentions)
Cyclin d1, by Beyotime (1 mentions)
42 protocols using image pro plus software v 6.0
1
Colony formation assay with xanthohumol
2
Evaluating Cytotoxic Effects of CTD
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Cells (2–3 × 103 cells/well) were seeded in 96-well plates, and after 24 h, they were treated with vehicle and different doses of CTD (2.5, 5 or 10 µM). Cell viability was determined by the MTT assay at 24, 48 and 72 h, respectively. For anchorage-independent cell growth assessment, cells (8 × 103 per well) were suspended in media containing 10% FBS for cell maintenance. After that, 0.3% agar with various concentrations of CTD was filled into a top layer upon a base layer of 0.5% agar containing the same corresponding doses of CTD in 6-well plates; three replicate wells were set for each concentration of the compound. After two weeks, the colonies were photographed using a microscope and calculated using the Image-Pro Plus software (v.6.0) program (Media Cybernetics, Rockville, MD, USA).
3
Soft Agar Colony Formation Assay
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Agar mix (0.5%) was prepared to form a base layer, which contained vehicle and HCPT (40, 80, and 160 nM) in the 6-well plate. Then, cells (8 × 103 cells/well) suspended in complete medium were added to 0.3% agar with vehicle, 40, 80, and 160 nM HCPT in a top layer over the base layer. Each of the agar plates were cultured and maintained in the incubator for 3 weeks and pictures were taken at the third week by microscope. Then, colonies were counted and summarized using the Image-Pro Plus software (v.6.0) program (Media Cybernetics, Silver Spring, MD, USA).
4
Quantifying Myocardial Infarct Size
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As described previously, the risk area and infarct size was calculated using a double-staining method with TCC and Evans blue stains (33 (link)). In brief, following perfusion, the heart was perfused with 3% Evans blue to indicate the area at risk (AAR). Then, the tissues were incubated with 2% TTC solution in the dark at 37°C for 15 min, and then were then stored in 4% paraformaldehyde overnight at 4°C to demarcate the infarct area. Areas with non-perfusion and blue staining were considered AAR, while areas with non-perfusion with TTC staining were labeled as the infarct area. The AAR and infarct area were calculated using Image-Pro Plus software V6.0 (Media Cybernetics, Inc., Rockville, MD, USA). The infarct size was measured as a percentage of the infarct area over the AAR.
5
Synthesis and Characterization of GNPs and SNPs
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GNPs ∼20 nm in size were prepared by the sodium citrate reduction of chloroauric acid [16 (link)], and SNPs ∼20 nm in size were prepared by the sodium borohydride reduction of silver nitrate [17 (link)]. The morphology of GNPs and SNPs was observed with transmission electron microscopy (TEM) (JEOL JEM-2100, Japan), and their sizes were determined with Image-Pro Plus software v6.0 (Media Cybernetics, Inc., USA). The concentrations of the GNPs and SNPs were measured using an inductive coupled plasma-optical emission spectrometer (Optima 5300DV, Perkin Elmer, USA).
6
Quantifying Fiber Density in Brain Regions
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Brain sections were analyzed using an Olympus Bx51 microscope (Optical Analysis Corporation, Nashua, NH, United States) at 400× magnification in conjunction with a Nikon DXM 1200 camera at 10× magnification. Image Pro Plus software v.6.0 (Media Cybernetics, Silver Springs, MD, United States) was used to superimpose a grid over the brain images. The number of fibers that definitively crossed the perimeter of the grid were counted and recorded. Counts were taken in the prelimbic cortex (PL) at Bregma +4.7, +3.7, and +2.7 mm; and anterior cingulate cortex (ACC) at Bregma +3.7, +2.5, and +0.7 mm anterior.
7
Immunofluorescence Staining and Microscopy
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Immunofluorescence staining was performed as previously described64 (link). Primary antibodies against anti-phospho-histone H2AX (Ser139/Tyr142) (1:100, Cell Signaling Technology, Cat 5438), calreticulin (1:400, Cell Signaling Technology, Cat 12238), and HMGB1 (1:400, Abcam, Cat ab18256) were used. Fluorescence-conjugated secondary antibodies (1:500, anti-rabbit, Dylight 488/594, Abbkine, Cat 23220/23430) and anti-fade fluorescence mounting medium with DAPI (ZSGB-BIO) were then used. The slides were examined with an FV1000 confocal microscope (Olympus) using FV10-ASW software v. 4.0 (Olympus). The fluorescence signals were analyzed using Image-Pro Plus software v. 6.0 (Media Cybernetics).
8
Histological Analysis of Nevus and Lymph Nodes
9
Quantitative Immunohistochemistry of Mouse Kidney
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Paraffin-embedded mouse kidney sections (3 μm) were prepared and performed immunohistochemical staining using routine protocols. The primary antibodies were indicated in Supplementary Table S2 . Some sections were stained with periodic acid-Schiff (PAS) (BA4080A, BASO). Images were photographed by Olympus BX53 microscope with EMCCD camera. At least ten glomeruli per mice in one section were analyzed to quantify positive area through the Image Pro plus software V6.0 (Media Cybernetics, Inc., Rockville, USA).
10
Quantifying HMGB1 in 4T1 Cells
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Different concentrations of COF samples were added to 4T1 cells and incubated overnight. Following various treatments, the 4T1 cells (2 × 103) were fixed with 4% paraformaldehyde, infiltrated with 0.5% Triton X-100 (BS084, Biosharp), blocked with 3% BSA (Biosharp BS114) in 0.1% triton/PBS for 1 hour, and incubated with anti-HMGB1 antibodies (1:400, ab18256, Abcam) at room temperature and Goat anti-Rabbit IgG DyLight 488 (1:200, A23220, Abbkine). The cells were then counterstained with DAPI (Beyotime Biotechnology, P0131), The slides were examined with an FV1000 confocal microscope (Olympus) using FV10-ASW software v. 4.0 (Olympus). The fluorescence signals were analyzed using Image-Pro Plus software v. 6.0 (Media Cybernetics).
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