Images of leaf layouts were obtained using a Nikon L110 digital camera. Photoshop and Illustrator (Adobe Creative Cloud; Adobe Systems Inc.) were used to prepare images and remove backgrounds of detached leaves for publication. A Nikon AZ100 microscope acquired micrographs of leaf stages. Image adjustments were made evenly within and consistently across figures and included background removal, as well as fine tuning of brightness, contrast and colour balance using Photoshop.
Creative cloud
Manufactured by Adobe
Sourced in United States
Creative Cloud is a collection of software and services developed by Adobe. It includes applications for creating and editing various types of media, such as graphics, photographs, videos, and web content. The core function of Creative Cloud is to provide users with a suite of tools for digital content creation and management.
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Lab products found in correlation
Az100 microscope, by Nikon (1 mentions)
Tissue freezing medium, by Leica (1 mentions)
Prolong gold antifade mountant, by Thermo Fisher Scientific (1 mentions)
Airyscan 2, by Zeiss (1 mentions)
Proteinase k, by Promega (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Tcs nt, by Leica (1 mentions)
Nis elements d3.2 imaging software, by Nikon (1 mentions)
Ds fi1 camera, by National Instruments (1 mentions)
Microphot fxa microscope, by National Instruments (1 mentions)
Shandon formal fixx 10 neutral buffered formalin, by Thermo Fisher Scientific (1 mentions)
Ds fi1, by Nikon (1 mentions)
Axiocam 506 mono camera, by Zeiss (1 mentions)
Plan apochromat 100 1.4 oil immersion objective, by Zeiss (1 mentions)
Prolong diamond antifade, by Thermo Fisher Scientific (1 mentions)
Filter set 49, by Zeiss (1 mentions)
Filter set 46, by Zeiss (1 mentions)
Axio observer z1 inverted microscope, by Zeiss (1 mentions)
12 protocols using creative cloud
1
Plant Leaf Imaging and Editing Protocol
2
Immunofluorescent detection of H3K9me3
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Embryos at 15-somite stage were fixed in 4% PFA (Sigma) for 2 h at RT, followed by washes in PBS. Embryos were then washed three times in 4% sucrose/PBS and allowed to equilibrate in 30% sucrose/PBS at 4°C for 3-5 h. Embryos were suspended in Tissue Freezing Medium (Leica) orientated in the sagittal plane and frozen with dry ice. Blocks were sectioned at 8 μm and slides were rehydrated in PBS, treated with −20°C pre-cooled acetone for 7 min at −20°C, washed three times with PBS and digested with Proteinase K (Promega) at a final concentration of 10 μg/mL for 3 min, washed 1x PBS and incubated in blocking buffer (10% Fetal Bovine Serum, 1% DMSO, 0.1% Tween20 in PBS) for 30 min. Primary antibody was diluted in blocking buffer and slides incubated overnight at 4°C. Slides were washed the following day and incubated with the appropriate AlexaFluor secondary antibodies (1:500), DAPI (0.5 μg/mL) and Phalloidin-TRITC (1:200) diluted in blocking buffer for 1 h at RT. Slides were washed, covered with glass coverslips with ProLong Gold Antifade Mountant (Thermofisher) and imaged at 63X with a LSM900 confocal with AiryScan2 (Zeiss). Images were viewed and processed in Imaris 9.3 (Bitplane) and Adobe Creative Cloud (Adobe). Primary antibody: anti-H3K9me3 abcam ab8898 at 1:500 (Chandra et al., 2012 (link)).
3
Microscopic Imaging of Lipid Staining
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A Nikon Microphot FXA microscope equipped with a DS-Fi1 camera and the NIS Elements D.32 imaging software (Nikon instruments Europe B.V., Badhoevedorp, The Netherlands) and a Leica multispectral confocal laser microscope (Leica TCS NT, Heidelberg, Germany) were used to examine the cryosections. Oil red O-stained lipids were detected under the confocal microscope using an excitation laser line at 543 nm (helium-neon). Optical sections (1.0 µm) were acquired, and images were produced using an 8-fold frame averaging a 1024 × 1024 pixel resolution. Representative images were presented using Adobe Creative Cloud (Adobe Inc., San Jose, CA, USA).
4
Histological Evaluation of Organ Samples
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The liver, lung, kidney and heart samples were first fixed in 10% buffered formalin (Shandon Formal-Fixx 10% neutral buffered formalin; Thermo Scientific) and then dehydrated and embedded in paraffin using the standard procedure. Subsequently, 5 µm histological sections were cut and stained with hematoxylin and eosin. Histology evaluation was performed using a light microscope (Ni/U; Nikon) equipped with a digital camera (DS-Fi1) and the NIS-Elements BR 4.60 imaging software (Nikon Instruments Europe B.V., Badhoevedorp, The Netherlands). Representative images are presented using Adobe Creative Cloud (Adobe Inc., San Jose, CA, USA).
5
Confocal Imaging of HA-YFP-PIF3 Proteins
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Confocal analysis of HA-YFP-PIF3 and HA-YFP-PIF3mTAD was performed as previously described with minor modifications 77 . Seedlings were fixed in 2% paraformaldehyde in PBS under vacuum for 15 min and then washed three times with 50 mM NH 4 Cl in PBS for 5 min three times. The seedlings were permeabilized with 0.2% Triton X-100 in PBS for 5 min, incubated with 300 nM DAPI in PBS for 10 min. The seedlings were washed three times with PBS for 5 min three times, and mounted with ProLong TM Diamond Antifade (ThermoFisher Scientific) on a slide. The slides were left to cure overnight in the dark, sealed with nail polish, and stored at 4°C until imaging. Nuclei of hypocotyl epidermal cells were imaged using a Zeiss Axio Observer Z1 inverted microscope equipped with a Plan-Apochromat 100×/1.4 oilimmersion objective and an Axiocam 506 mono camera (Carl Zeiss, Jena, Germany).
Fluorescence was detected with the following Zeiss filter sets: YFP, exciter 500/25 nm/nm, emitter 535/40 nm/nm (Zeiss Filter Set 46); DAPI, exciter 365 nm, emitter 445/50nm/nm (Zeiss Filter Set 49). Images were collected using Zeiss ZEN software and processed using Adobe Creative Cloud (Adobe, San Jose, CA).
Fluorescence was detected with the following Zeiss filter sets: YFP, exciter 500/25 nm/nm, emitter 535/40 nm/nm (Zeiss Filter Set 46); DAPI, exciter 365 nm, emitter 445/50nm/nm (Zeiss Filter Set 49). Images were collected using Zeiss ZEN software and processed using Adobe Creative Cloud (Adobe, San Jose, CA).
6
Blinded Statistical Analysis Protocol
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The experimenters were blinded to grouping/genotype information during data collection and analyses. Sample sizes and number of independent experiments were estimated by power analyses using an R script (“pwr” package on CRAN) that takes pre-specified effect size, type I and II errors as input arguments. Shapiro–Wilk test and F test were first employed to test normality and equal variance. All data that passed normality/equal variance tests were reported as mean ± SEM (standard error of the mean). For normal-distributed/equal variance data, Student's t-test or multiple t-tests were used. Statistical analyses and graphing were performed using GraphPad Prism 8.0, Microsoft Excel, and MATLAB. P-Value < 0.05 was considered statistically significant for all tests. Figures were prepared using Adobe Creative Cloud.
7
Assembling Figures Using Creative Cloud
8
Comprehensive Imaging Protocol for Cellular Analysis
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Figure plates were created using Adobe Illustrator and Adobe Photoshop in Adobe Creative Cloud. cLSM scans were prepared and imaged using Imaris × 64 9.7.2 (developed by Bitplane).
9
Vessel Segmentation from Fundus Images
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Original FA images (2392 × 2048 pixel) were processed, traced into binary (black/white) images, and analyzed at various zoom levels using 2019 Photoshop Adobe Creative Cloud on a 15.6‐inch HP laptop at a resolution of 1366 × 768 pixel. A color fundus image was used to define the difference between arteries and veins that were separated from each other according to physiological vascular branching rules.14 , 15 , 16
10
Acoustic Analysis of Jet Overflight
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A large passenger jet aeroplane in a steady overflight was recorded with a sampling rate of 44,100 Hz, stereo channel selection, and a 16-bit depth on Adobe Creative Cloud, Audition® CC 2017 (v.10.1), Adobe Creative Cloud, and presented at a level of 80 dBLAeq. The 15-second “overflight” was preceded and followed by silence. Noise reduction techniques were applied and wind noise removed from the recording which was trimmed to 15 seconds. Onset and offset were smoothed using a spline curve. The frequency spectrum and relative levels were not altered from the original recording. Peak clipping was not apparent. Spectral analysis of the noise showed a dominance of middle frequency components (750–1500 Hz).
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