Image pro plus 5

Manufactured by Media Cybernetics

Sourced in United States, Japan, Germany, United Kingdom, China

Image-Pro Plus 5.0 is an image analysis software that provides a suite of tools for capturing, processing, analyzing, and managing digital images. The software offers a range of functions, including image acquisition, enhancement, measurement, and quantification, as well as the ability to create custom analysis workflows.

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Lab products found in correlation

Bx51 microscope, by Olympus (5 mentions) Fluorescence microscope, by Olympus (5 mentions) Tcs sp5, by Leica (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (4 mentions) Lsm 710, by Zeiss (4 mentions) Fast green, by Merck Group (4 mentions) Odyssey infrared imaging system, by LI COR (3 mentions) Nissl staining, by Beyotime (3 mentions) Hoechst 33342, by Thermo Fisher Scientific (3 mentions) 4 6 diamidino 2 phenylindole (dapi), by Merck Group (3 mentions) Dapi fluoromount g, by Southern Biotech (3 mentions) Axio observer z1, by Zeiss (3 mentions) 3 3 diaminobenzidine (dab), by Merck Group (3 mentions) Triphenyl tetrazolium chloride (ttc), by Merck Group (3 mentions) Fv3000, by Olympus (3 mentions) Goat anti brn3a rgc marker antibody, by Santa Cruz Biotechnology (3 mentions) Dexamethasone (dex), by Merck Group (3 mentions) β glycerophosphate, by Merck Group (3 mentions) Safranin o fast green, by Merck Group (3 mentions) Ix71 microscope, by Olympus (2 mentions)

443 protocols using image pro plus 5

Quantifying Cardiac Fibrosis and Apoptosis

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After 24–48 h in 4% paraformaldehyde, the blocks were dehydrated and embedded in paraffin, cut into 4 μm slices, heated overnight in a 60°C incubator, and then dewaxed and stained with H&E and Masson dye. One slice was chosen from each rat and was analyzed under a microscope. Masson staining for the presence of interstitial collagen fiber accumulation was a marker of cardiac fibrosis. The ratio of interstitial fibrosis to the total left ventricular area was calculated from 20 randomly selected microscopic fields in five individual sections per heart using a camera attached to an Leica DM2000 microscope, with images further analyzed by Image-Pro Plus 5.1 (Media Cybernetics, Silver Spring, MD), excluding coronary vessels and perivascular regions.
Apoptosis in cardiac tissue was determined with the DeadEnd Fluorometric TUNEL System (Promega), which catalytically incorporates fluorescein-12-dUTP at DNA strand breaks as previously described [16 (link)]. All sections were counterstained with DAPI (Molecular Probes) at a final concentration of 2 μM. Images were viewed with epifluorescence microscopy (Leica TCS SP5) within 24 hours and analyzed with Image-Pro Plus 5.1 Software (Media Cybernetics, Silver Spring, MD).

Li Y., Fang J., Hua Y., Wang C., Mu D, & Zhou K. (2014). The Study of Fetal Rat Model of Intra-Amniotic Isoproterenol Injection Induced Heart Dysfunction and Phenotypic Switch of Contractile Proteins. BioMed Research International, 2014, 360687.

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Histochemical Analysis of Muscle Fibers

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Histochemical analyses were performed with ATPase staining using the method of Brooke and Kaiser [22 (link)] with slight modifications. Transverse serial sections of 10 µm in thickness were cut from entire blocks (1.0 × 1.0 × 1.5 cm) with a cryostat microtome (HM525, Microm GmbH, Walldorf, Germany) at −20 °C. Sections were subsequently used for histochemical analysis of myosin adenosine triphosphatase (mATPase) following alkaline (pH 10.70) preincubation. An image analysis system (Image-Pro^® plus 5.1, Media Cybernetics Inc., Rockville, MD, USA) was used to examine the stained sections. The muscle fibers were classified as fiber types I, IIA and IIB according to the nomenclature of Brooke and Kaiser. Approximately 500 fibers per sample were counted to analyze the muscle fiber characteristics with Image-Pro Plus software (Image-Pro^® plus 5.1, Media Cybernetics Inc., Rockville, MD, USA), including the fiber number percentage (FNP) and fiber area percentages (FAP). FNP shows the ratio of the counted fiber number of each fiber type to the total counted fiber number. FAP was the ratio of a total cross-sectional area of each fiber type to total fiber area measured.

Li Y., Ma Q., Shi X., Yuan W., Liu G, & Wang C. (2022). Comparative Transcriptome Analysis of Slow-Twitch and Fast-Twitch Muscles in Dezhou Donkeys. Genes, 13(9), 1610.

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Osteogenic and Adipogenic Differentiation of PDLSCs

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For osteogenesis, the PDLSCs were plated at a density of 5×10³ cm² in 24-well plates and cultured for 3 days. The cells were then incubated with a-MEM, supplemented with 10% FBS, 50 µM L-ascorbic acid 2-phosphate (Sigma-Aldrich), 10 mM β-glycerophosphate (Sigma-Aldrich) and 100 nM dexamethasone (Sigma-Aldrich), for 2 weeks to induce mineral formation. The cells were fixed with 70% ethanol for 15 min and stained with 2% alizarin red (pH 4.0; Sigma-Aldrich) for 15 min. The nodule area was measured quantitatively using an image analysis system (Image-Pro Plus 5.0; Media Cybernetics, Inc., Baltimore, MD, USA).
For adipogenesis, the PDLSCs were plated at a density of 5×10³ cm² in 24-well plates and were cultured for 3 days. The cells were then incubated with a-MEM, supplemented with 10% FBS, 0.5 mM methylisobutylxantine, 0.5 µM hydrocortisone and 60 µM indomethacin (Sigma-Aldrich). The cells were cultured for an additional 21 days. The adipogenic cultures were fixed in 70% ethanol for 15 min and stained with 2% fresh Oil Red O solution (Sigma-Aldrich) for 15 min. The lipid area was measured quantitatively using an image analysis system (Image-Pro Plus 5.0; Media Cybernetics, Inc.).

ZHENG W., WANG S., WANG J, & JIN F. (2015). Periodontitis promotes the proliferation and suppresses the differentiation potential of human periodontal ligament stem cells. International Journal of Molecular Medicine, 36(4), 915-922.

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Placental ROS Quantification using DHE

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Dihydroethidium probes were used to reflect the level of ROS in the placenta. Dihydroethidium could cross cell membranes at the presence of ROS and stain nuclei bright red by intercalating with the DNA [50 (link)]. At first, the frozen placenta was sliced into 10 μm cross-sections which were then incubated with 5 μmol dihydroethidium (37 °C, 15 min). A Nikon 2000S fluorescence microscope (Nikon, Melville, NY, USA) and the Image-pro Plus 5.0 (Media Cybernetics, Inc., Rockville, MD, USA) were applied respectively to observe and calculate the level of ROS (mean fluorescence intensity per unit area calculated by Image-pro Plus 5.0).

Yu M., Chen L., Peng Z., Wang D., Song Y., Wang H., Yao P., Yan H., Nüssler A.K., Liu L, & Yang W. (2017). Embryotoxicity Caused by DON-Induced Oxidative Stress Mediated by Nrf2/HO-1 Pathway. Toxins, 9(6), 188.

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Retinal Layer and RGC Quantification

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Thicknesses of retinal layers (outer segments + inner segments, outer nuclear layer, outer plexiform layer, inner nuclear layer, inner plexiform layer, retinal ganglion cell layer) were measured for treated and non-treated eyes of WES (n = 4) and Sham (n = 3) rats from 20× magnification images of retinal cross sections obtained via a phase contrast microscope (Leica DMLB, Leica Inc., Buffalo Grove, IL) using an image analysis program (Image-Pro Plus 5.0; Media Cybernetics, Warrendale, PA). Retinal regions spanning 2.5 mm superiorly and inferiorly from the optic nerve head were measured. Each 2.5 mm region was subdivided into five 0.5 mm sections and designated “ F” or “ S” 1–5 for inferior and superior, respectively. Thicknesses for each retinal layer were compared between Sham and WES groups at each location examined. Additionally, thicknesses across all locations examined for each retinal layer were averaged within experimental group and then compared.
RGC nuclei were quantified using an image analysis program (Image-Pro Plus 5.0; Media Cybernetics, Warrendale, PA). RGC counts were averaged in each of the ten regions in both WES (n = 5) and Sham (n = 9) eyes. Additionally, summed RGC counts of superior and inferior regions 1–4 were compared between experimental groups. All nuclei in the RGC layer were counted which included RGCs and any displaced amacrine cell nuclei.

Hanif A.M., Kim M.K., Thomas J.G., Ciavatta V.T., Chrenek M., Hetling J.R, & Pardue M.T. (2016). Whole-eye electrical stimulation therapy preserves visual function and structure in P23H-1 rats. Experimental eye research, 149, 75-83.

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Western Blot Analysis of Cell Proteins

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Whole-cell proteins were extracted and subjected to standard western blot analysis as described previously [41 (link)]. Antibodies against MYOCD (SAB4200539, Sigma, Massachusetts, USA), ACTA2(ab7817, Abcam, Massachusetts, USA), SM22α (ab14106, Abcam, Massachusetts, USA), PCNA (sc-56, Santa Cruz, California, United States), OPN (sc21742, Santa Cruz, California, United States), LC3 (NB100-2220, Novus Biologicals, Colorado, USA), P62 (610833,BD Transduction Laboratories, USA), Beclin1 (ab207612, Abcam, Massachusetts, USA) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (ab8245, Abcam, Massachusetts, USA) were used to probe for the target proteins. Secondary antibodies, including IRDye^®800CW anti-rabbit secondary antibody and IRDye^®680 anti-mouse secondary antibody, were purchased from Li-COR (Lincoln, Michigan, USA). Signals were detected by an Odyssey™ Infrared Imaging System (LI-COR, Michigan, USA), and the bands were quantified by Image-Pro Plus 5.1 software (MEDIA CYBERNETICS, Maryland, USA).

Shi D., Ding J., Xie S., Huang L., Zhang H., Chen X., Ren X., Zhou S., He H., Ma W., Zhang T, & Wang N. (2022). Myocardin/microRNA-30a/Beclin1 signaling controls the phenotypic modulation of vascular smooth muscle cells by regulating autophagy. Cell Death & Disease, 13(2), 121.

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Immunohistochemical Characterization of Vascular Smooth Muscle

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All of the samples were fixed in 4% paraformaldehyde and embedded in paraffin. Then 5 μm serial sections were generated and evaluated by hematoxylin and eosin (H&E) and immunohistochemistry staining 9 (link),25 (link). Anti-Myh 11 (mouse monoclonal, 1:300 dilution, Novus Biologicals) and anti-α-SMA antibody (mouse monoclonal, 1:500 dilution, Abcam) were used to verify the smooth muscle origin. Anti-CD31 (mouse monoclonal, 1:250 dilution, Novus Biologicals) and anti-CD34 (rat monoclonal, 1:250 dilution, Thermo Fisher Scientific) antibodies were used to verify the vascular origin. The sections were heated in 10 mM Tris-HCl buffer (pH 8.8) for antigen retrieval and the samples treated with primary antibodies were incubated overnight at 4℃. Then the sections were incubated with appropriate secondary antibodies for 1 h at room temperature, followed by subsequent linking to horseradish peroxidase and substrate/chromogen reactions using an immunoperoxidase secondary detection kit (Millipore, Billerica, MA, USA). Negative controls without primary antibodies were prepared to rule out non-specific labeling. The slides were observed using an Axioplan 2 microscope (Zeiss). The positive staining was quantified using Image-Pro Plus 5.1 software (Media Cybernetics, Inc., Rockville, MD, USA) in 8 different fields for each tissue sample.

Guo H.L., Peng X.F., Bao X.Q., Wang L., Jia Z.M., Huang Y.C., Zhou J.M., Xie H, & Chen F. (2020). Bladder reconstruction using autologous smooth muscle cell sheets grafted on a pre-vascularized capsule. Theranostics, 10(23), 10378-10393.

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Histological Analysis of Oral Tissue Response

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Tissue biopsies were performed one day after bleaching, as it is the optimal time to evaluate any reaction. Rabbits were euthanized, and teeth, gingiva, tongue, buccal mucosa, and hard and soft palates were harvested for histological analysis. All biopsied tissues were placed in different labeled vials containing 4% paraformaldehyde fixative solution at room temperature for at least 48 h. Teeth were decalcified in neutral 10% ethylenediaminetetraacetic acid (EDTA) for four weeks, and the EDTA solution was changed every day to accelerate demineralization. The fixed tissues were embedded in paraffin under vacuum. Serial sections of 4 μm thickness were cut using a rotary microtome, mounted on glass slides, and subjected to hematoxylin and eosin (H&E) and Masson’s trichrome (MST) staining. H&E staining was performed to evaluate epidermal or dermal changes and morphological changes of tissue; MST, the three-color staining in histology, was used to detect collagen fibers. The stained tissues were examined microscopically, in a blinded manner, using a light microscope (CKX41, Olympus, Tokyo, Japan) adapted with a digital camera (Pixel link PL-B686 CU, Canada), at a magnification of ×100, ×200, and ×400. Images were loaded into a computer and processed with Image-Pro Plus 5.1 software (Media cybernetics Inc., Washington, DC, USA).

Nam S.H., Choi B.B, & Kim G.C. (2021). The Whitening Effect and Histological Safety of Nonthermal Atmospheric Plasma Inducing Tooth Bleaching. International Journal of Environmental Research and Public Health, 18(9), 4714.

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Multiparametric Immunofluorescence Analysis of Skin Wound Healing

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CD29 and CD90 were used to label MSCs; CK14 and CK19 for different epithelial subsets; and S-100A4 for fibroblasts. The rabbits were euthanized at the indicated time points. The wounds were soaked in 30% sucrose in PBS overnight at 4°C and then embedded in Tissue-Tek O.C.T (Sakura Finetek USA, Inc., Torrance, CA, USA). Serial 8 mm thick sections were cut at −20°C and placed on poly-L-lysine-coated microscopic slide. Non-specific labeling was blocked with 10% normal goat serum for 30 min at room temperature. The sections were incubated with primary antibodies against CD29 (bs-3973R; Bioss), CD90 (ab225; Abcam), CK14 (ab77684; Abcam), CK19 (ab84632; Abcam), and S-100A4 (bs-3759R; Bioss) at 4°C overnight. Omitting primary antibodies was performed as control staining. After several washes, the secondary antibodies (ab150077; Abcam) were applied followed by incubation for 60 min at room temperature. An Olympus BX51 microscope equipped with fluorescence and a CCD camera was used to capture the fluorescence images. The IOD was evaluated using Image-Pro Plus 5.1 software (Media Cybernetics, Inc.).

Liu J., Hu F., Tang J., Tang S., Xia K., Wu S., Yin C., Wang S., He Q., Xie H, & Zhou J. (2017). Homemade-device-induced negative pressure promotes wound healing more efficiently than VSD-induced positive pressure by regulating inflammation, proliferation and remodeling. International Journal of Molecular Medicine, 39(4), 879-888.

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Western Blot Analysis of Metabolic Regulators

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Western blot analysis was performed as previously reported17 (link). The antibodies previously used for Western blots were the anti-human TCF7L2 antibody (1:800; 13838-1-AP, Proteintech, USA), anti-human PEPCK antibody (1:1000; 14892-1-AP; Proteintech, USA), anti-human GLUT2 antibody (1:800; 20436-1-AP; Proteintech, USA), anti-human IRS2 antibody (1:1000; 20702-1-AP; Proteintech, USA), anti-human pAKT antibody (1:1000; 60072-1-Ig; Proteintech, USA), anti-human AKT antibody (1:1000; 10176-2-AP; Proteintech, USA), anti-human pGSK antibody (1:1000; 14850-1-AP; Proteintech, USA), anti-human GSK antibody (1:1000; 22104-1-AP; Proteintech, USA), anti-human pERK1/2 antibody (1:1000; 3441-100; BioVision, USA), anti-human ERK1/2 antibody (1:1000; 16443-1-AP; Proteintech, USA), and anti-human GAPDH antibody (1:1000; 10494-1-AP; Proteintech, USA). GAPDH was used as control. The results were quantified by the ImagePro Plus 5.1 software (Media Cybernetics, Inc., USA).

Dong F., Ling Q., Ye D., Zhang Z., Shu J., Chen G., Fei Y, & Li C. (2016). TCF7L2 involvement in estradiol- and progesterone-modulated islet and hepatic glucose homeostasis. Scientific Reports, 6, 24859.

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