Following surgical resection, the entire gland was embedded and axially step-sectioned (Pucar et al., 2005 (link)). Whole mount prostatectomies were fixed overnight in 10% neutral buffered formalin (Fisher). The samples were dehydrated in graded alcohol, embedded in paraffin and 5 μm sections used for analysis. Sections were stained with haemotoxylin and eosin (H & E) and used for immunohistochemical analysis performed on an automated Ventana machine (Discovery, Ventana Medical Systems Inc.).
Ventana machine
Manufactured by Roche
Sourced in United States
The Ventana machine is a piece of laboratory equipment designed for automated slide staining and processing. It is used to prepare tissue samples for microscopic analysis in medical and research settings. The core function of the Ventana machine is to automate the process of applying reagents and performing staining procedures on histological and cytological specimens, enabling efficient and consistent sample preparation.
Automatically generated - may contain errors
Lab products found in correlation
Neutral buffered formalin, by Thermo Fisher Scientific (1 mentions)
Hrp conjugated goat anti rabbit antibody, by Agilent Technologies (1 mentions)
Nis elements software, by Nikon (1 mentions)
Anti mouse cd45 antibody clone 30 f11, by BD (1 mentions)
Ndp view, by Hamamatsu Photonics (1 mentions)
Au2700 autoanalyser, by Olympus (1 mentions)
5 protocols using ventana machine
1
Detailed Prostate Histopathology Workflow
2
Liver Pathology Characterization and Fibrosis Assessment
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Liver pathology was characterized from H&E stained sections based on the semi-quantitative NAFLD activity score (NAS) which evaluates the degree of steatosis, lobular infiltrates and hepatocytes ballooning [42 (link)]. Liver fibrosis was assessed by Sirius red coloration, and the stained area calculated from the total liver surface using NIS-Elements software (Nikon). Immunolocalisation of IL-33 was performed using a primary goat IgG anti-mouse-IL-33 antibody (R&D Systems) and a secondary HRP-conjugated rabbit anti-goat antibody (Dako, USA) with hematoxylin counterstaining in a Ventana machine (Ventana Medical Systems, Inc. USA). Serum biochemical analyses of transaminases levels (AST/ALT) were performed as described previously [9 (link)].
3
Acute Toxicity Evaluation of Reduced Graphene Oxide
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This research was approved by the Institutional Ethics Committees of the First Affiliated Hospital of Chongqing Medical University (Approval notice: # 2018-77) and conformed to the tenets of the Declaration of Helsinki. 40 C57BL/6 mice aged 6–8 weeks were randomly divided into four groups (n = 10, one-half male and one-half female) and given a single oral dose of 0, 100, 200, or 1,000 mg/kg of RGO. The LD50 assay was performed with the maximum dose (1,000 mg/kg). Twenty mice were randomly divided into control and treated groups (n = 10). The treated group received oral RGO, 1,000 mg/kg, once. The mental state and activity of the mice were observed daily and their weight measured once a week. All mice were sacrificed 14 days after drug administration. Collectable organs including brain, heart, lungs, stomach, pancreas, liver, kidneys, ovaries, uterus, bowel, and spleen were harvested right after sacrifice. Samples were prepared with formalin and embedded with paraffin using a Ventana machine.
4
Liver Inflammation Markers and Immune Cell Profiling
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The histopathological (hematoxylin and eosin (H&E) staining) and level of liver transaminases (ALT/AST) in serum were performed as described earlier. Immunolocalisation of IL-33 was performed by immunohistochemical staining using primary antibody anti-mouse-IL-33 (goat IgG, R&D Systems). Leucocytes were stained with anti-mouse CD45 antibody (clone 30-F11, BD Pharmingen), macrophages with anti-mouse F4/80 antibody (clone BM8, eBioscience), and neutrophils, after antigen retrieval step (citrate buffer pH 6), with anti-mouse Ly6G antibody (clone 1A8, BD Bioscience). Then, secondary HRP-conjugated dedicated specific antibodies (Dako, USA) were used followed by hematoxylin counterstaining in a Ventana machine (Ventana Medical Systems Inc., USA). The counting of stained positive cells was carried in at least 3 different microscopic fields of 1 mm2 surface area by tissues on at least 3 different tissues by conditions by using image analysis software (NDP, view, Hamamatsu).
5
Histopathological Analysis of Liver
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The histopathological analysis of liver was performed using haematoxylin & eosin–safran staining. Levels of serum alanine and aspartate transaminases (ALT and AST) were assessed according to the IFCC primary reference procedures using Olympus AU2700 Autoanalyser® (Olympus Optical, Tokyo, Japan). Immunolocalization of IL‐33 or CXCL10 was performed by histochemical staining using primary goat anti‐mouse‐IL‐33 and anti‐CXCL10 (R&D Systems, Minneapolis, MN) and secondary horseradish peroxidase‐conjugated rabbit anti‐goat antibody for IL‐33 (Dako, Markham, Ont., Canada) and OmniMap anti‐Rabbit‐horseradish peroxidase (RUO) for CXCL‐10 followed by haematoxylin counterstaining in a Ventana machine (Ventana Medical Systems, Inc. Tucson, AZ), as previously described.36 Counting of necrotic areas and inflammatory foci was carried out on liver areas of 15–30 mm2 using the NDP.2 view image analysis software (Hamamatsu Photonics K.K., Japan).
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