Cells and frozen renal tissue sections were fixed with 4% paraformaldehyde in PBS for 10 min at room temperature, washed with PBS, and permeabilized with 0.3% Triton X-100 for 10 min. After blocking in 1% BSA for 30 min, the cells were immunolabeled with primary antibodies (anti-Kim-1 at 1:150 dilution; anti-TNF-α at 1:100 dilution; anti-dsDNA at 1:400 dilution; anti-TOM20 at 1:100 dilution; or anti-TFAM at 1:50 dilution) overnight at 4 °C followed by incubation with FITC- or TRITC-conjugated secondary antibody (1:200) for 1 h at 37 °C. Nuclei were visualized by staining with DAPI for 5 min at room temperature. Digital images of the sections were captured using a fluorescence microscope (Nikon, N-STORM & A1, Tokyo, Japan), and the Pearson's correlation coefficient between the TFAM and dsDNA results was analyzed using Image-Pro Plus.
Image pro plus
Manufactured by Nikon
Sourced in Japan, United States, Australia
Image-Pro Plus is a comprehensive image analysis software developed by Nikon. It provides advanced tools for image processing, measurement, and analysis. The software offers a wide range of features for quantifying and interpreting digital images from various sources, including microscopes and other imaging equipment.
Automatically generated - may contain errors
Lab products found in correlation
Model 2100, by A-M Systems (2 mentions)
Smz25, by Nikon (2 mentions)
A1r confocal microscope, by Nikon (2 mentions)
Eclipse 80i, by Nikon (2 mentions)
Labchart 5, by ADInstruments (2 mentions)
N storm a1, by Nikon (1 mentions)
Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions)
Anti ki67 primary antibody, by Abcam (1 mentions)
Eclipse ts100, by Nikon (1 mentions)
Formalin, by Merck Group (1 mentions)
Coolpix, by Nikon (1 mentions)
Dxm1200, by Nikon (1 mentions)
In situ cell death detection kit, by Roche (1 mentions)
Chamber slide, by Thermo Fisher Scientific (1 mentions)
Lysotracker red dnd 99, by Thermo Fisher Scientific (1 mentions)
Anti ang 2, by Abcam (1 mentions)
Eclipse 80i microscope, by Nikon (1 mentions)
G1003, by Wuhan Servicebio Technology (1 mentions)
Eclipse e100, by Nikon (1 mentions)
Matrigel, by BD (1 mentions)
29 protocols using image pro plus
1
Immunofluorescence Analysis of Renal Proteins
2
Immunohistochemistry of Jejunum Tissues
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Jejunum tissues were formalin fixed and paraffin embedded, and 3 mm sections were dewaxed in the following order: xylene I for 15min, xylene II for 15 min, anhydrous ethanol for 15 min, 85% ethanol for 5 min, 75% ethanol for 5 min, distilled water for 5 min.Then the antigen was repaired with EDTA repair solution (pH: 8.0–9.0) and hydrophobic histochemical pen was used to block the hydrosphere. After the slides were sealed in 3%–5% BSA for 30–60 min primary antibody was added and incubated overnight at 4°C, followed by respective secondary antibodies conjugated with fluorescence.
IEC-6 cells were treated with 4% paraformaldehyde for 15 min, then treated with 0.1% Triton/PBS at 4 °C and permeabilized for 10–20 min according to the expression position of the target protein. Then IEC-6 cells were incubated with 3–5% BSA for 1 h followed by primary antibody overnight at 4 °C. On the second day, the jejunum sections and IEC-6 cells were washed with PBS for 3 times, 5min/per time, and then incubated with corresponding fluorescence secondary antibody for 1 h. After washed with PBS 3 times, 5min/per time, DAPI was used to stain the nuclei. Nikon A1R confocal microscope was used for evaluation, Image J and Image-Pro Plus software was used for Image analysis.
IEC-6 cells were treated with 4% paraformaldehyde for 15 min, then treated with 0.1% Triton/PBS at 4 °C and permeabilized for 10–20 min according to the expression position of the target protein. Then IEC-6 cells were incubated with 3–5% BSA for 1 h followed by primary antibody overnight at 4 °C. On the second day, the jejunum sections and IEC-6 cells were washed with PBS for 3 times, 5min/per time, and then incubated with corresponding fluorescence secondary antibody for 1 h. After washed with PBS 3 times, 5min/per time, DAPI was used to stain the nuclei. Nikon A1R confocal microscope was used for evaluation, Image J and Image-Pro Plus software was used for Image analysis.
3
Cell Proliferation and Viability Assay with TG003
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To count live cells, 3 × 105 cells were seeded into six-well plates containing 2 ml of cell culture medium and treated with 1, 10 and 50 µM TG003. Every 24 h cells were trypsinised and counted in a haemocytometer (Neubauer chamber). Trypan blue (0.4% w/v) was used as an exclusion stain.
To measure cell proliferation, 3 × 105 cells were seeded into six-well plates containing coverslips and serum-starved in 4 ml of DMEM for eight hours, then replaced with full medium. Cells were treated with 1 µM, 10 µM and 50 µM TG003. After 48 h incubation, media was removed and cells were washed twice. Washed cells were fixed in 4% (w/v) paraformaldehyde (PFA) for ten minutes and permeabilized in 0.25% (v/v) triton in PBS.
Fixed cells were blocked in 10% FBS-PBS for one hour and incubated overnight at 4 °C with 1:1000 anti-Ki67 primary antibody (Abcam, Cambridge, UK). Cells were washed three times in PBS before incubation for one hour at room temperature with secondary antibody, AlexaFluor 488 goat anti-rabbit (Molecular Probes), diluted 1:500, followed by washing. Cells were then counterstained with DAPI for four minutes and further washed twice before mounting. Images were obtained with an Image Pro Plus (TE 300 Nikon Japan) microscope and the percentage proliferation calculated with ImageJ software.
To measure cell proliferation, 3 × 105 cells were seeded into six-well plates containing coverslips and serum-starved in 4 ml of DMEM for eight hours, then replaced with full medium. Cells were treated with 1 µM, 10 µM and 50 µM TG003. After 48 h incubation, media was removed and cells were washed twice. Washed cells were fixed in 4% (w/v) paraformaldehyde (PFA) for ten minutes and permeabilized in 0.25% (v/v) triton in PBS.
Fixed cells were blocked in 10% FBS-PBS for one hour and incubated overnight at 4 °C with 1:1000 anti-Ki67 primary antibody (Abcam, Cambridge, UK). Cells were washed three times in PBS before incubation for one hour at room temperature with secondary antibody, AlexaFluor 488 goat anti-rabbit (Molecular Probes), diluted 1:500, followed by washing. Cells were then counterstained with DAPI for four minutes and further washed twice before mounting. Images were obtained with an Image Pro Plus (TE 300 Nikon Japan) microscope and the percentage proliferation calculated with ImageJ software.
4
Bone Microarchitecture and Cell Analysis
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Harvested sternum samples were fixed, embedded, and sliced. For microarchitecture, the sections were stained with toluidine blue, the micrographs of the bone sections were taken using a microscope (Nikon Eclipse TS100; Nikon Corporation, Tokyo, Japan), and an image analyzer (Image Pro-Plus, Rockville, MD) at a magnification of 20×. Bone static histomorphometry parameters were performed by a blinded examiner using Weibel Grid technique, which includes the trabecular bone volume/tissue volume (BV/TV ), trabecular number (Tb.N ), Tb.Th , and spacing (Tb.Sp ). For osteoblasts or osteoclasts detection, the sections were stained with alkaline phosphatase (ALP ) or tartrate-resistant acid phosphatase (TRAP ) detecting kit (Sigma-Aldrich, St. Louis, MO), respectively. The ALP positive staining represents osteoblast, and TRAP-positive staining represents osteoclasts. Osteoblast and osteoclast number per bone surface (N.Ob/BS and N.Oc/BS ) were counted on the external surfaces of the bone using the surgical defect as the field of view. A single operator at 2 separate time points performed the quantification using Image Pro-Plus.
5
Quantifying Liver Metastases Burden
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At the endpoint laparotomy was performed and the abdominal cavity examined for indications of macroscopic extra- hepatic metastases; paying close attention to the splenic bed. The liver was carefully excised and harvested. Immediately following excision liver weights were recorded. Organs were then fixed in formalin (10 %) (Sigma Aldrich, Castle Hill, NSW, Australia) for 24 h and then transferred to 70 % ethanol. Images of the whole liver were taken to examine the tumor distribution and burden load. The liver was then transversely sliced into sections of 1.5 mm thickness using a tissue fractionator. For large livers (Saline control and OXi4503 treated) every second slice was sampled for analysis. For smaller livers (Sunitinib and Sunitinib/OXi4503 treated tumors) every slice was taken for analysis due to small number of sections. Liver slices were placed on a clear plastic plate; a digital camera (Nikon Coolpix5000, E500) was used to capture the images and these were analysed using image analysis software (Image Pro Plus, Perth, Australia).
Tumors were visualised as distinctive white/cream coloured areas against the red/brown liver tissue. Each tumor outline was traced using image analysis software to determine the area occupied by the tumor. This stereology technique was used to determine the number of tumor nodules, tumor volume and the percentage of liver metastases.
Tumors were visualised as distinctive white/cream coloured areas against the red/brown liver tissue. Each tumor outline was traced using image analysis software to determine the area occupied by the tumor. This stereology technique was used to determine the number of tumor nodules, tumor volume and the percentage of liver metastases.
6
Cerebral Infarct Quantification
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After 24h of reperfusion, mice were sacrificed and their brains were immediately dissected out, sliced, and stained with 2% 2,3,5-triphenyltetrazolium chloride (TTC) for 15 min at 37°C. Then brain sections were fixed in 4% formaldehyde overnight. Slices images were taken with a digital camera (Nikon, Coolpix) and analyzed using image analysis software (Image-Pro Plus, Version 6.0). Infarct volume was measured by the equation:
Infarct volume (%) = Infarct volume/ Total volume of slice × 100%.
Infarct volume (%) = Infarct volume/ Total volume of slice × 100%.
7
Evaluating Oligodendrocyte Progenitor Cell Viability
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The viability of OPCs was evaluated by terminal deoxinucleotidyl transferase-mediated dUTP-fluorescein nick end labeling (TUNEL) staining using an In Situ Cell Death Detection Kit (Roche), according to the manufacturer’s protocol. Fluorescence images were captured using a fluorescence microscope (DXM1200, Nikon) and the proportion of TUNEL+/NG2+ cells of the total cells was quantified using Image-Pro Plus.
8
Dextran and BSA Internalization in BMDCs
9
Quantifying Cell Migration and Morphology
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A series of images was taken with an image analyser immediately before field exposure and during 24 hours (1–5, 10 and 24 h) after field exposure [4] (link), [7] (link). A minimum of ten fields per slide were taken, two slides were analyzed, and three to five replicates per period of time were evaluated. Individual frames were recorded and analyzed using light microscope Nikon Eclipse 80i and Image-Pro Plus software.
Mean migration rate was quantified over 5 h [4] (link), [28] (link), and elongation and orientation were quantified over 24 h.
Mean migration rate was quantified over 5 h [4] (link), [28] (link), and elongation and orientation were quantified over 24 h.
10
Immunohistochemical Analysis of Angiotensin II in Rat Adipose Tissue
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The fat tissues from 6- and 16-week-old rat offspring were collected and incubated in 4% paraformaldehyde, embedded in paraffin, and sectioned. The sections were incubated with anti-Ang II (1∶200, Abcam, UK) for 2 h at 37°C and then with goat anti-rabbit IgG conjugate (Santa Cruz, USA) for 30 min at 37°C. After each incubation, the slides were washed 3 times with Tris-buffered saline with Tween 20 (TBST) for 5 min each. The slides were then counterstained with Mayer's hematoxylin for 15 s. When observed and photographed under the microscope, positive regions appear brown and yellow. Light micrographs were captured with a color video camera (Nikon Microscope E-100, Japan) and analyzed with image analysis software (Image Pro Plus, USA).
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