The following apparatus was used: Las4000 Mini Imager (GE Company, U.S.A.); Image Pro Plus 6.0 Image Analysis System (Media Cybernetics Company, U.S.A.); Reward R407 small animal ventilator (Shenzhen Reward Life Technology Co., Ltd., China).
Image pro plus 6.0 image analysis system
Manufactured by Media Cybernetics
Sourced in United States
Image-Pro Plus 6.0 is an image analysis software system that enables users to capture, enhance, measure, count, and analyze digital images. It provides a suite of tools for a variety of imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Bx53 microscope, by Olympus (1 mentions)
D700 digital camera, by Nikon (1 mentions)
Imagequant las 4000 mini imager, by GE Healthcare (1 mentions)
Caspase 1, by Thermo Fisher Scientific (1 mentions)
Nlrp3, by Abcam (1 mentions)
A0208, by Beyotime (1 mentions)
Bx53m, by Olympus (1 mentions)
Erk1 2, by Abcam (1 mentions)
Western blot electrophoresis apparatus, by Bio-Rad (1 mentions)
Ht7800, by Hitachi (1 mentions)
Anti foxp3, by Cell Signaling Technology (1 mentions)
Anti cd3, by Proteintech (1 mentions)
Anti p akt, by Proteintech (1 mentions)
Anti akt, by Proteintech (1 mentions)
Ab25022, by Abcam (1 mentions)
Affinipure goat anti rabbit igg, by ZSGB-BIO (1 mentions)
Zli 9031, by ZSGB-BIO (1 mentions)
Rhodamine conjugated goat anti rabbit igg, by Proteintech (1 mentions)
17 protocols using image pro plus 6.0 image analysis system
1
Small Animal Imaging Protocol
2
Immunostaining of KS and Angioma Tissues
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Twelve KS tissues, one normal skin tissue, and six non-KS angioma tissues were collected. All the samples were formalin-fixed, paraffin-embedded, and immunostained with anti-AR or anti-LANA antibodies at a ratio of 1:200 as previously described [30 (link)]. The results were processed using Image-Pro plus 6.0 image analysis system (Media Cybernetics, MD, USA).
3
Quantifying Atrial Fibrosis via Trichrome Staining
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Left atrial tissue was fixed in 10% phosphate-buffered formalin, embedded in paraffin, sliced into 4 µm serial sections, and subjected to pathological examination following hematoxylin-eosin (H&E) staining and Masson's trichrome staining. Photomicrographs were obtained using an Olympus BX53 microscope (Olympus, Tokyo, Japan). Masson's trichrome staining was used to evaluate atrial interstitial fibrosis. The collagen fibers are marked with blue, while the cardiomyocytes are marked with red. Semiquantitative analysis of the proportion of collagen fibers was conducted using the Image-Pro Plus 6.0 image analysis system (Media Cybernetics, Maryland, USA). The proportion of fibrous tissue area was calculated by the following equation: collagen fiber area/total view area 100%.
4
Quantifying Brain Infarct Volume in Rats
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Rats were sacrificed by deeply anaesthetized with 5% isoflurane and euthanized by cervical dislocation. Brain tissues were kept at −20 °C for 30 min after ice cold saline washing. Coronal sections (3 mm) of the frozen brains were cut with the sharp blades and stained with 2% 2, 3, 5-triphenyltetrazolium chloride (TTC, dissolved in PBS solution, pH 7.4) at 37 °C for 30 min in the dark. Sections were reversed every 5 min to keep uniform coloring of the sections. Viable tissues were stained in deep red while the infarcts remain unstained. TTC-stained sections were photographed with a digital camera after being fixed with 4% paraformaldehyde. Finally, images were captured by a Nikon D700 digital camera (Nikon, Japan).
The infarct volume of each section was calculated using Image-Pro Plus 6.0 image analysis system (Media Cybernetics, USA). The infarct area (IS) was calculated as previously described using the following equation35 (link): IS = (1 − S1/S2)*100% (S1: ipsilateral hemisphere non-infarcted area; S2: contralateral hemispher area)
The infarct volume of each section was calculated using Image-Pro Plus 6.0 image analysis system (Media Cybernetics, USA). The infarct area (IS) was calculated as previously described using the following equation35 (link): IS = (1 − S1/S2)*100% (S1: ipsilateral hemisphere non-infarcted area; S2: contralateral hemispher area)
5
Small Animal Ventilation and Imaging
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The apparatus used was as follows: Reward R407 small animal ventilator (Shenzhen Reward Life Science & Technology Co., Ltd., China); Image Quant LAS4000 mini imager (GE, USA); Image Pro Plus 6.0 image analysis system (Media cybernetics Co., USA); and Radox blood gas analyzer ABL80 (RADIOMETER, Denmark).
6
Immunohistochemical Analysis of NLRP3, Caspase-1, IL-1β, and IL-18
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Paraffin sections were incubated in 3% hydrogen peroxide at 25 °C for 5-10 min to inhibit endogenous peroxidase activity. Primary antibodies (NLRP3: Abcam, Shanghai, China; caspase-1: Thermo Scientific, Waltham, MA, United States; and IL-1β and IL-18: Boster Biotechnology, Wuhan, China) were added and incubated at 4 °C overnight, washed with phosphate-buffered saline (PBS), and incubated with the appropriate secondary antibody at 20-37 °C for 10-30 min. Haematoxylin and eosin staining was performed, as described previously[40 (link)]. The optical density and area of each image were measured using an Image-Pro Plus 6.0 Image Analysis System (Media Cybernetics Inc., Bethesda, MD, United States), and the mean density was calculated.
7
Histological Analysis of Spinal Cord
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After the electrophysiological experiment, the tissue sections were analyzed according to the above operation, followed by phosphate-buffered saline (PBS) hydration, and sealing with skimmed milk. Hematoxylin and eosin (H&E) staining was performed for histological analysis. The cytoplasm stained red with strong reflectivity, and the color changed with SNI, electrical stimulation, and medication. At room temperature, anti-CB1 receptor antibodies (Abclonal, A1447) were added and incubated for 2 h, followed by incubation with secondary antibodies (A0208,1:1000, Beyotime, Shanghai, China) for 1 h. After three PBS washes, the film was sealed and observed under a light microscope.
The Mshot High Definition Color Pathological Graphic Report Management System (Mshot MD50) was used to acquire the images of lumbar 2 and 3 segments of the spinal cord of neonatal, juvenile, and adult rats, respectively. Under a 200× light microscope, the superficial region of the spinal cord's posterior horn from rat sections was selected for H&E staining. Densitometric analysis was performed using Image ProPlus 6.0 image analysis system (Media Cybernetics). The optical density (OD) reflected the positive staining intensity of the tissue; the higher the OD, the stronger the staining. All OD measurements were conducted under the same optical conditions.
The Mshot High Definition Color Pathological Graphic Report Management System (Mshot MD50) was used to acquire the images of lumbar 2 and 3 segments of the spinal cord of neonatal, juvenile, and adult rats, respectively. Under a 200× light microscope, the superficial region of the spinal cord's posterior horn from rat sections was selected for H&E staining. Densitometric analysis was performed using Image ProPlus 6.0 image analysis system (Media Cybernetics). The optical density (OD) reflected the positive staining intensity of the tissue; the higher the OD, the stronger the staining. All OD measurements were conducted under the same optical conditions.
8
Apoptosis Quantification in Myocardial Tissue
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Myocardial tissue was fixed with 10% paraformaldehyde, dehydrated, and embedded in paraffin, and conventional sections were 4 µm. An anti-rabbit/mouse universal immunohistochemistry kit was used to process paraffin sections according to standard procedures. Reagent 1 (TdT) and reagent 2 (fluorescein-labeled dUTP) mixed solution (1 : 9, freshly prepared) (50–100 µL) was added to the cells and incubated. Then, 2-(4-amidinophenyl)-6-indolecarbamidine dihydrochloride (DAPI) (50–100 µL) was added and incubated at room temperature in the dark for 10 min and washed thrice in PBS in the dark for 5 min each. The slides were mounted with an antifluorescence quencher, and apoptotic cells were counted randomly in each section under six high-power field (×400) fluorescence microscopes. Image-Pro Plus 6.0 Image Analysis System (Media Cybernetics,US) was used to perform optical density analysis and record the integrated optical density (IOD) value.
9
Immunohistochemical Analysis of CXCL16 and ERK1/2
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The clinical tissues were provided by Shandong Provincial Hospital for IHC and were then fixed in 10% formaldehyde for 24 h, immersed in 75% ethanol for 2 h followed by tissue dehydration: 80% ethanol 20 min, 90% ethanol 20 min, 95% ethanol 20 min, 100% ethanol I 10 min, 100% ethanol II 10 min, xylene I 10 min, xylene II 10 min, xylene III 10 min, paraffin I 40 min, paraffin wax II 1 h, Paraffin III 1 h and embedded in paraffin. The tissue was sectioned at 5 µm and attached to slides. After blocking endogenous peroxides and non-specific proteins, the tissues sections were incubated with primary antibodies, CXCL16 (cat. no. ab119350, Abcam) and ERK1/2 (cat. no. ab17942, Abcam) at 4°C overnight. The following day, the sections were washed three times and were incubated with HRP-goat-anti-rabbit antibody (1:10,000; cat. no. ab205718, Abcam) at 37°C for 30 min. The tissues sections were then stained with diaminobenzidine for 3 min and the nuclei were counterstained with hematoxylin for 2 min at room temperature. The CXCL16 and ERK1/2 antibodies were used at a dilution of 1:200. IHC images of each tissues were randomly obtained using a light microscope (BX53M; Olympus Corporation), then were analyzed using the Image-Pro Plus 6.0 image analysis system (Media Cybernetics, Inc.).
10
Molecular Mechanisms of Puerarin in Ischemic Stroke
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The following drugs, reagents, and instruments were used in the study (Supplementary Table 1 ): puerarin injection (Xi'an Hanfeng Pharmaceutical Co., Ltd.; specification: 2 mL: 100 mg, H20033784); MCAO monofilaments (Changsha Maiyue Biotechnology Co., Ltd., China, M8500); triphenyltetrazolium chloride (TTC) staining reagent (Shanghai Jizhi Biochemical Technology Co., Ltd., China, T54930-25 g); antiSIRT1 (ab28170); antiHIF-1α (Ab1003), antiVEGF (ab10284), antisynaptophysin (SYN) (ab93133), antipostsynaptic density protein (PSD)-95 (ab2723), secondary antibody goat antirabbit immunoglobulin G (ab7085), and antiGAPDH antibody (ab8245) (purchased from Abcam, UK); interleukin (IL)-1β enzyme-linked immunosorbent assay (ELISA) kit (Beijing Soleibao Technology Co., Ltd., China, Cat. No.: SEKG-0002-96Time); IL-6 ELISA kit (Nanjing Saihongrui Biotechnology Co., Ltd., China, Cat. No.: HEA079Hu03); intercellular adhesion molecule (ICAM)-1 ELISA kit (Nanjing Sunberga Biotechnology Co., Ltd., China, Cat. No.: SBJ-H0008); Western blot electrophoresis apparatus (Bio-Rad, USA); TDZ4B-WS low-speed centrifuge (Shanghai Luxiangyi Co., Ltd., China); Image-ProPlus 6.0 image analysis system (Media Cybernetics Co., Ltd., USA); and transmission electron microscope (Huison Biotechnology [Shanghai] Co., Ltd., China).
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