The lateral view of a whole brain of D. sargus (Fig. 2A) was shot with a digital camera (Sony Handycam DCR-SR52). The representative photomicrographs of the coronal sections (Fig. 2A, a-d) were captured with a digital camera (Olympus Sc30) mounted on Olympus SZX7 zoom stereo microscope, and using the Cell-P software (Olympus, Germany). An Olympus BX61 microscope (Olympus, Germany), coupled with a digital camera (Olympus DP) and running the Cell-P software (Olympus, Germany) was used to obtain the photomicrographs of typical cell morphology (Fig. 2B) at a final magnification of 1000×.
Cellp software
Manufactured by Olympus
Sourced in Germany, Japan
CellP software is a digital imaging platform that enables the acquisition, processing, and analysis of microscopic cell samples. It provides a comprehensive set of tools for image capture, segmentation, and quantification of cellular features.
Automatically generated - may contain errors
Lab products found in correlation
Bx61 microscope, by Olympus (5 mentions)
Xm10 camera, by Olympus (3 mentions)
Ix81 fluorescence microscope, by Olympus (3 mentions)
Synthetic mountant, by Thermo Fisher Scientific (3 mentions)
Superfrost microscope slide, by Avantor (3 mentions)
Mca1477, by Bio-Rad (3 mentions)
Ax70 microscope, by Olympus (2 mentions)
Orca er digital camera, by Olympus (2 mentions)
Ix70 fluorescence microscope, by Olympus (2 mentions)
Handycam dcr sr52, by Sony (2 mentions)
Szx7 zoom stereo microscope, by Olympus (2 mentions)
Bx41 microscope, by Olympus (2 mentions)
Streptavidin alexa 488, by Thermo Fisher Scientific (2 mentions)
Cy3 conjugated goat anti rabbit antibody, by Jackson ImmunoResearch (2 mentions)
Mowiol, by Merck Group (2 mentions)
Bx51m reflection epifluorescence microscope, by Olympus (1 mentions)
Abc elite kit, by Vector Laboratories (1 mentions)
Donkey anti rabbit biotin, by Jackson ImmunoResearch (1 mentions)
Orca digital camera, by Hamamatsu Photonics (1 mentions)
M3 84, by BD (1 mentions)
49 protocols using cellp software
1
Brain Imaging of Diplodus sargus
2
Detailed Brain Imaging Protocols
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The representative photomicrographs of the coronal sections (Fig. 2 A-D) were captured with a digital camera (Olympus Sc30) mounted on Olympus SZX7 zoom stereo microscope, and using the Cell-P software (Olympus, Germany). The photograph of lateral view of a whole brain of D. sargus (Fig. 2 A'-D') was shot with a digital camera (Sony Handycam DCR-SR52). The photomicrographs of stained sections in Fig. 3A andB were shot with a digital camera (Olympus DP) mounted on an Olympus BX61 microscope (Olympus, Germany) with a 4x (UPlan SApO, N.A. 0.16) objective or 100x (UPlan SApO, N.A. 1.4) oil objective, using the Cell-P software (Olympus, Germany).
3
Quantitative Analysis of Neuronal Fluorescence
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Photographs of the stained motor cortex and thalamus sections from three rostrocaudal anatomical levels from bregma (AP: −1.70, −2.06, and − 2.30) were taken at ×10 magnification. We used Cell P software (Olympus Soft Imaging Solutions, Münster, Germany) from an Olympus DP70 digital camera connected to an Olympus AX 70 microscope (Olympus, Zoeterwoude, The Netherlands). In the images of the area of interest, fluorescent density was measured using ImageJ software (version 1.52; NIH, Bethesda, USA). The average value of three sections was used for statistical analysis in each subject.
4
Platelet Activation Assay Protocol
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PRP samples were perfused through the device in the presence or absence of TRAP and ASA for 30 minutes. Afterward, the samples were incubated with fluorescein isothiocyanate-conjugated PAC-1 antibodies (BD Biosciences, Oxford, UK) for 30 minutes at room temperature in the dark. PAC-1 recognizes an epitope on the activated glycoprotein GPIIb/IIIa complex of activated platelets in live cells. The samples were then fixed with 90% ethanol, rinsed in PBS, and mounted on a glass slide with mounting medium.
Photomicrographs were captured using a fast, high-resolution XM10 camera (Soft Imaging System GmbH, Münster, Germany) mounted on an Olympus BX51M reflection epifluorescence microscope. Images were processed using Cell-P software (Soft Imaging System).
Photomicrographs were captured using a fast, high-resolution XM10 camera (Soft Imaging System GmbH, Münster, Germany) mounted on an Olympus BX51M reflection epifluorescence microscope. Images were processed using Cell-P software (Soft Imaging System).
5
Quantifying c-Fos Expression in Brain Regions
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Photographs of the stained motor cortex and thalamus sections from three rostrocaudal anatomical levels from bregma (AP: −0.58, −0.94, and −1.22) were taken at 10X magnification. We used Cell P software (Olympus Soft Imaging Solutions, Münster, Germany) from an Olympus DP70 digital camera connected to an Olympus AX 70 microscope (Olympus, Zoeterwoude, The Netherlands). In the images of the area of interest, the number of c-Fos–positive cells was counted using ImageJ software [version 1.52; National Institutes of Health (NIH), Bethesda, USA]. Cells immunopositive for c-Fos were counted manually, and the mean number of cells was corrected for surface area and expressed as cells per square millimeter. A cell was regarded positive when the intensity of the cell staining was significantly higher than the surrounding background. The average value of three sections was used for statistical analysis in each subject. For the subthalamic area (the infusion site), a digital photograph was taken at one anatomical bregma (−2.06), and all c-Fos–positive cells within 1 mm2 of the injection site were counted.
6
Immunohistochemical Analysis of c-Fos Expression
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Sections were incubated for three and two nights in Study 1 and Study 2, respectively, with anti-c-Fos primary antibody (1:2000; rabbit polyclonal; Abcam ab209794). After washing with Tris-buffered saline (TBS) and TBS-Triton X-100 (TBS-T), sections were incubated with secondary antibody (1:800, donkey anti-rabbit biotin; Jackson Immunoresearch Laboratories Inc., Westgrove, PA, USA) for 1 h. This was followed by repeated washing and incubation with avidin–biotin peroxidase complex (1:800, Elite ABC kit, Vectastain, Burlingame, CA, USA) for 2 h. The DAB combined with NiCl2 intensification was used to visualize the staining, and then dehydrated and coverslipped with Pertex (Histolab Products ab, Göteborg, Sweden).
Photomicrographs were also taken with Cell P software (Olympus Soft Imaging Solutions, Münster, Germany) using an AX-70 microscope (Olympus, Zoeterwoude, The Netherlands) with 10× magnification. Cells immunopositive for c-Fos were counted manually after reducing the background by an appropriate threshold level, and then divided by the surface area of interest (cells/mm2). In Study 1, the classical auditory regions were analysed, including PAC, the medial part of MGB and the central part of IC and DCN. For Study 2, c-Fos expression was investigated in PAC, the central part of the IC, CA1 of the hippocampus, the dentate gyrus, the basolateral amygdala, the TRN and the DCN.
Photomicrographs were also taken with Cell P software (Olympus Soft Imaging Solutions, Münster, Germany) using an AX-70 microscope (Olympus, Zoeterwoude, The Netherlands) with 10× magnification. Cells immunopositive for c-Fos were counted manually after reducing the background by an appropriate threshold level, and then divided by the surface area of interest (cells/mm2). In Study 1, the classical auditory regions were analysed, including PAC, the medial part of MGB and the central part of IC and DCN. For Study 2, c-Fos expression was investigated in PAC, the central part of the IC, CA1 of the hippocampus, the dentate gyrus, the basolateral amygdala, the TRN and the DCN.
7
Quantitative Analysis of Microglia Activation
8
Time-lapse Wound Closure Imaging
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Time lapse images were captured at 15-minutes intervals on a Leica DM IRB phasecontrast inverted microscope (Leica; Milton Keynes, UK) in a chamber maintained at 36 ± 1°C and 5%CO 2 atmosphere. The images were collected with a cooled Hamamatsu ORCA digital camera (Hamamatsu Photonics, Welwyn Garden City, UK) connected to a computer running Cell^P software (Olympus, London, UK) over 48-hours (until complete wound closure). For quantitative analysis of time lapse serial images ImageJ software (Schneider et al. 2012 ) was used.
9
CREB and NFκB Activation in HUVECs
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For CREB experiments, HUVECs were treated with 0.6 μM of rMASP-1, 400 μM of CoCl 2 , or both for 2 h with or without 6 μM of C1INH. A second plate of HUVECs was placed in a hypoxic (1% O 2 ) incubator with or without 0.6 μM of rMASP-1 treatment for 2 h. In another experimental setup, HUVECs/HUAECs were treated with varying concentrations of rMASP-1 (0.06 μM, 0.2 μM, 0.6 μM, 2 μM), 400 μM of CoCl 2 , or the combination of each dose of rMASP-1 with CoCl 2 for 2h. For NFκB experiments, HUVECs were treated with 400 μM of CoCl 2 for 2 h with/without 0.6 μM of rMASP-1 in the last 1h. We used 1 ng/ml of IL-1β as a positive control in both CREB and NFκB experiments. Cells were then fixed in Methanol-Acetone and stained with rabbit-anti-human phospho-CREB (pCREB, 1:200) or rabbit-anti-human NFκB (1:250) followed by Alexa Fluor568-conjugated goat anti-rabbit (1:500) IgG and Hoechst 33342 (1:50000). For NFκB experiments, additional Alexa Fluor568-conjugated wheat germ agglutinin (WGA, 1:200) staining was added after for 20 min. Olympus IX-81 fluorescence microscope and an Olympus XM-10 camera were used to take photos. Photos were analyzed using CellP software (Olympus). Nuclear mean red fluorescence (pCREB) or the difference between cytoplasmic and nuclear mean red fluorescence (NFκB) were calculated.
10
Neutrophil Adhesion Assay on ICAM-1
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