For imaging of flat-mounted embryos after in situ hybridisation, an AxioZoom V16 stereomicroscope (Zeiss) equipped with an Axiocam 506-Mono and a colour digital camera were used. Immunostained embryos were imaged with Zeiss LSM 800 or 880 with Airyscan confocal microscopes. For live imaging, embryos were aligned on heptane glue coated coverslips and submersed in a thin layer of halocarbon oil. Bright-field live imaging was performed using an AxioZoom V16 stereomicroscope, while fluorescence live imaging was performed with confocal microscopes. Image stacks were processed in Fiji (Schindelin et al., 2012 (link)) and Helicon Focus (HeliconSoft). Image brightness and intensity was adjusted in Corel PhotoPaint X5 (CorelDraw) and Fiji.
Photopaint x5
Manufactured by Corel
Sourced in Canada
PhotoPaint X5 is a versatile image editing software that provides a range of tools and features for manipulating digital images. Its core function is to allow users to edit, enhance, and create visual content with precision and efficiency.
Automatically generated - may contain errors
Lab products found in correlation
Axio zoom v16 stereomicroscope, by Zeiss (1 mentions)
Axiocam 506 mono, by Zeiss (1 mentions)
Tsa fluorescein, by PerkinElmer (1 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
Axiocam 506, by Zeiss (1 mentions)
Ds fi2, by Nikon (1 mentions)
Leica dmi 4000b inverted microscope, by Leica Microsystems (1 mentions)
Leica application suite, by Leica Microsystems (1 mentions)
E600 light microscope, by Nikon (1 mentions)
Eos 30d digital camera, by Canon (1 mentions)
Photo paint x3, by Corel (1 mentions)
B 100ap, by Analytik Jena (1 mentions)
Eos 40d digital camera, by Canon (1 mentions)
6 protocols using photopaint x5
1
Imaging and Analysis of Flat-Mounted Embryos
2
In situ hybridization with fluorescence detection
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RNA in situ hybridisation was carried out as described in [71 (link)], with the following minor modifications: Proteinase K treatment and post-fixations steps in the original protocol were omitted, and prior to hybridization, the probes were heated to 80 °C for 5 min and immediately put on ice before adding to the pre-hybridization buffer. Fluorescent in situ hybridization was performed following [40 ]. Tyramide Signal Amplification (TSA) was performed with TSA kits from PerkinElmer (TSA Fluorescein and TSA Cyanine). Post hybridisation, nuclear staining was achieved by incubating embryos in 1 μg/ml 4–6-diamidino-2-phenylindol (DAPI) in PBS with 0.1% Tween-20 for 15 min. Embryos were mounted in glycerol on Poly-L-lysine (Sigma-Aldrich) coated coverslips, where the germband tissue attaches making it easier to remove the yolk before imaging. Images were taken with an AxioZoom V16 stereomicroscope (Zeiss) equipped with an Axiocam 506 mono and colour digital camera. Brightness and intensity of the pictures were adjusted in Corel PhotoPaint X5 (CorelDraw).
3
Microscopy and Drawing of Gonopods
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Measurements and photographs were made with a Nikon DS-Fi2 camera mounted on a Nikon SMZ25 stereo microscope, using the NIS-Elements Microscope Imaging Software with an Extended Depth of Focus (EDF) and edited in Adobe Photoshop CS6.
Gonopods drawings were performed with pencil and then black liners using a Olympus SZ-61 stereo microscope, sketches were scanned with a CanoScan Lide 60 scanner and then edited with Corel Photo-Paint X5 software. Figure plates were assembled in Adobe Photoshop CS6.
Gonopods drawings were performed with pencil and then black liners using a Olympus SZ-61 stereo microscope, sketches were scanned with a CanoScan Lide 60 scanner and then edited with Corel Photo-Paint X5 software. Figure plates were assembled in Adobe Photoshop CS6.
4
Automated Spheroid Morphology Analysis
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The morphology of the spheroids was visualized by a Leica DMI4000 B inverted microscope (Leica Microsystems, Wetzlar, Germany) with 5× magnification objective. For assessment of ATN-RNA effects in 3D cultures on spheroid morphology we applied author’s spheroid-edge detection method. In brief, for each ATN-RNA concentration at experimental endpoint (72 h) high magnification microscopy images were taken with a Leica Application Suite (Leica Microsystems, Wetzlar, Germany) with a 1 ms exposure time. Central micro tumours were cropped into the final images for spheroid-contour processing. The size of each cropped image in all experiments was 567 x 567 pixels (24 Bit Color RGB), with resolution of 96 dpi. In next step, the contour of spheroids was identified, followed by Contour Tool in Corel® PHOTO-PAINT X5 (Corel Corporation, Ottawa, ON, Canada). The size of the spheroids was analysed using the analysis tools in Photoshop CS5 software (Adobe Systems, San Jose, CA, USA).
5
Photomicrographic Documentation of Reticulinasus salahi
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The images of the larvae specimens of Reticulinasus salahi from Oman (Fig. 1 a, b) were taken by an Olympus XC30 digital camera installed on the Nikon E600 light microscope, where the bright-field and interference contrasts (NomarskyDIC) were applied. For processing the photos, analysis Docu v. 5.1 and Corel Photopaint X5 were employed. The photos of an adult female of R. salahi from Iran (Fig. 2 a, b) were made using a Canon EOS 30D digital camera and Canon MP-E 65/2.8 Macro lens in multiple layers, stacked using Helicon Focus and edited in Corel Photopaint X3.![]()
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Photomicrograph parts of capitulum of Reticulinasus salahi larvae from locality Ain Sahnawt, Oman.
Slightly engorged nymph of Reticulinasus salahi from locality Bishapur, cave at the Sâsân spring, Iran, originally identified as Ornithodoros sp. in Benda et al. (2012 : 530).
6
Transient expression assay in Nicotiana benthamiana
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Leaves of the transgenic line 16c of N. benthamiana [30 (link)] were co-infiltrated “on the spot” [58 (link)] with A. tumefaciens (pGV2260) transformed for expression of different proteins. For infiltration, A. tumefaciens cultures for expression of mgfp4 [36 (link)], HCpro, HCproY345A;L351A, VPgY89A;L94A, and Tiso4E were diluted with infiltration medium to a final OD600 = 0.5 and for co-infiltration, the cultures were combined at equal ratios. If any of these constructs were not needed for a given experiment, the corresponding A. tumefaciens strain was replaced with a strain expressing GUS [55 (link)]. GFP accumulation in the infiltrated leaves was detected using a hand-held UV-lamp (B-100 AP; UVP). Images were taken with an EOS 40D digital camera (Canon, Amsterdam, The Netherlands) and processed using Corel PHOTO-PAINT X5 (Corel Corporation, Ottawa, Canada).
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