Wild type zebrafish larvae (AB strain) were purchased from Shanghai FishBio Co., Ltd. Larvae of AB strain were cultured in medium (0.16 mmol/L MgSO4, 0.33 mmol/L CaCl2, 0.17 mmol/L KCl, 5 mmol/L NaCl, and 10 ppm methylene blue) under isothermal conditions at 28.5°C. On 9 dpf (days post-fertilization), the zebrafish larvae were stained with alizarin red. First, zebrafish larvae were immobilized in 4% polyformaldehyde for 2.5 h and dehydrated in 50% ethanol for 20 min. Then, zebrafish larvae were bleached with 1.5% H2O2 in 1% KOH for 25 min to remove the pigment and washed 3 times with PBST. Next, zebrafish larvae were stained with 0.01% alizarin red staining in 0.7% KOH for 4 h. Finally, the zebrafish larvae were decolorized in different proportions of 0.5% KOH and glycerin (3:1, 1:1, 1:3) for 6–8 h. The experimental protocols for quantifying larval skull mineralization were similar to those previously published (Wu et al., 2021 (link)). Zebrafish were placed on a slide covered with glycerin. Stereomicroscope was used to photograph ventral view and lateral view of zebrafish. The mineralization area and integrated optical density (IOD) of skull alizarin red staining were analyzed using Image-Pro Plus (IPP, Media Cybernetics, United States).
Image pro plus ipp
Manufactured by Media Cybernetics
Sourced in United States
Image Pro Plus (IPP) is a comprehensive image analysis software designed for advanced microscopy and imaging applications. It provides a suite of tools for image acquisition, processing, measurement, and analysis. IPP supports a wide range of image file formats and integrates with a variety of imaging hardware and devices.
Automatically generated - may contain errors
Lab products found in correlation
Inverted microscope, by Zeiss (2 mentions)
Ab134936, by Abcam (1 mentions)
Ab207299, by Abcam (1 mentions)
Active β catenin, by Merck Group (1 mentions)
Ultrasensitive s p kit, by Maixin Group (1 mentions)
Ivis lumina series 3, by PerkinElmer (1 mentions)
Expose mouse and rabbit specific hrp dab detection ihc kit, by Abcam (1 mentions)
9 protocols using image pro plus ipp
1
Alizarin Red Staining of Zebrafish Larvae
2
Immunohistochemical Analysis of Biomarkers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Standard streptavidin-peroxidase immunohisto-chemistry procedures were performed following the manufacturer's instructions with an UltraSensitive TM SP Kit (Maixin-Bio, Fujian, China) as previously described [36 (link)]. Primary antibodies against PLCD1 (ab134936; Abcam), MMP7 (ab207299; Abcam), pERK1/2 (#4370; Cell Signaling Technology), or active-β-catenin (05-665; Millipore) were employed, and PBS was used as a negative control. All immunohistochemical images were subjected to mean optical density (OD) measurement using Image Pro Plus (IPP, version 6.0; Media Cybernetics, Silver Spring, MD, USA).
3
Quantifying Osteogenic Differentiation of HBMSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Calcium deposition of HBMSCs was determined using Alizarin red S staining after 21 days of osteogenic induction. HBMSCs were fixed for 40 min in 4% paraformaldehyde fluid at room temperature, washed twice with PBS, and then stained with 40 mm Alizarin red S staining at pH 4.2 for 20 min with gentle agitation. Mineralized nodules were visualized using an inverted microscope (Carl Zeiss AG, Oberkochen, Germany) and measured by image‐pro plus (ipp ; Media Cybernetics, Manassas, VA, USA) analysis. Ten images were captured for each group, and the mean percentage was calculated.
4
Nanoparticle Tumor Imaging and Biodistribution
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
1,1′-Dioctadecyl-3,3,3′,3′-tetramethylindotricarbocyanine (DiR)-labeled PTX-A10-3.2-PLGA NBs and PTX-PLGA NBs 0.2 mL (20 mg/mL) were injected into LNCaP tumor-bearing nude mice via the tail vein. The changes in fluorescence intensity in xenograft tumors at the time intervals of 0, 2, 4, 8, 12, and 24 h were assessed in vivo using a small animal living fluorescence imaging apparatus (IVIS Lumina Series III; PerkinElmer Inc., Waltham, MA, USA). DiI-labeled PTX-A10-3.2-PLGA NBs and PTX-PLGA NBs 0.2 mL (20 mg/mL) were also injected into LNCaP tumor-bearing nude mice via the tail vein. The nude mice were euthanized, and the tumor tissue, liver, spleen, and kidney were removed and brought to the pathology department immediately to obtain cryosections. All pieces of the organ tissues were stained using DAPI to visualize the cell nucleus. The samples were washed three times with normal saline after incubation for 5 min at room temperature. The organ tissues were treated with antifade mounting medium, and then the fluorescence was observed by LCSM. We measured the integrated optical density (IOD) of each image using Image Pro Plus (IPP; Media Cybernetics, Inc, Rockville, MD, USA) 6.0 for quantitative analysis.
5
Immunohistochemical Analysis of PCNA and Caspase-3
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The sections were pretreated with xylene and then incubated with primary antibody against rat proliferating cell nuclear antigen (PCNA) or Caspas-3 (Santa Cruz Biotechnology, Inc.), at 4 °C overnight. Next, sections were incubated with complement, horseradish peroxidase (HRP) conjugate, and diazoaminobenzene (DAB), orderly (Abcam EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit). Next, sections were counterstained with hematoxylin. Images were captured by a microscopy (Olympus). The images of immunostained sections were analyzed using Image Pro-Plus (IPP) software (Media Cybernetics, Silver Spring, MD, USA) to calculate the density mean, area sum, and integrated optical density (IOD) of positive expression. The PCNA and Caspase-3 levels were denoted by Average optical density (AOD). AOD = IOD(sum)/Area(sum).
6
Renal Histomorphometry of Minipigs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tissues from the same locations in the kidneys of the minipigs were collected (including the cortex and medulla) and fixed in 4% paraformaldehyde and 85% ethanol for conventional paraffin sectioning at a thickness of 6 μm. The structure of the renal tissue was observed by hematoxylin-eosin (HE) and periodic acid–Schiff (PAS) staining. The thicknesses of the cortices and medullas were measured to calculate the cortex/medulla ratio using an H-7500 (Japan) electron microscope and Image-Pro Plus (IPP; Media Cybernetics Corporation, Maryland, USA). Under the 40X eyepiece of an H-7500 (Japan) electron microscope, five random fields were selected to calculate the areas of the glomeruli and the tubular unit. Frozen sections were stained with Oil Red O to examine the accumulation of lipids. The sections were observed using an H-7500 electron microscope.
7
Immunohistochemical Detection of sLex
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were seeded on glass coverslips and then fixed with PBS containing 4% formaldehyde for 10 min and permeabilized with 0.2% Triton X-100 in PBS for 10 min at room temperature. After washing with PBS, endogenous peroxidase activity was suppressed by 3% H2O2 and blocked with goat serum or 5% bovine serum albumin (BSA). Diluted primary monoclonal rabbit antibodies against sLeX (dilute 1:100) were added and incubated at 4°C overnight. As secondary reagents, biotin-labeled anti-IgG and an avidin–biotin HRP complex were used Zhongshan jinqiao (Beijing, China) followed by staining with chromogen diaminobenzidine (Zhongshan) until a brown color developed. Slides were counterstained with Mayer’s hematoxylin and differentiated in a solution containing 1% hydrochloric acid and 99% ethanol. Sections were dehydrated, and a transparent cover slip was added to enable observation by microscopy. All the immunohistochemical photographs were analyzed using Image-Pro Plus (IPP; Media Cybernetics, Inc., Rockville, MD, USA).
8
Histological Analysis of HIF-2α, LOXL2, MMP-9, E-cadherin, and Snail
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For histological examination, 4‐μm sections were stained with H&E and IHC using standard protocol. Primary antibodies used: anti–HIF‐2α (1:100), anti–LOXL2 (1:100), anti–MMP‐9 (1:200), anti–E‐cadherin (1:200) and anti–Snail (1:200). Data were collected from an average of 10 randomly selected areas for each section by using Image Pro‐Plus (IPP) (Media Cybernetics, Silver Spring, MD, USA).
9
Cartilage Matrix Metabolism during OA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To further clarify cartilage matrix metabolism during OA progression, glycosaminoglycans (GAG) and matrix metalloproteinase-3 ( MMP-3) expression in cartilage were detected by immunohistochemistry. Briefly, tissue sections were routinely deparaffinized, rehydrated, and repaired in complex phosphoesterasum for 10 min at 37 °C, and then incubated overnight at 4 °C with anti-rabbit GAG (1:200; Boster Corporation, Wuhan, China) and MMP-3 (1:150; Boster Corporation, Wuhan, China) antibodies. The remaining procedures were adapted from the PV-6001 Two-Step IHC Detection Reagent illustration (ZSGB-BIO Corporation, Beijing, China); the brown color was developed using DAB (ZSGB-BIO Corporation, Beijing, China) and the sections were counterstained with hematoxylin (Baso Corporation, Zhuhai, China).
All sections were semi-quantitatively analyzed using Image Pro Plus (IPP; Media Cybernetics Inc., Rockville, MD, USA) version 6.0 software, and the integrated optical density (IOD) was measured from the images at 200× magnification.
All sections were semi-quantitatively analyzed using Image Pro Plus (IPP; Media Cybernetics Inc., Rockville, MD, USA) version 6.0 software, and the integrated optical density (IOD) was measured from the images at 200× magnification.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!