Frozen tumor sections were sequentially fixed with cold acetone, acetone plus chloroform (1:1), and acetone. Paraffin-embedded sections were deparaffinized and heated in antigen retrieval buffer. Tissue sections were blocked with 3% H2O2 in distilled water for 20 min and then in blocking buffer (5% normal horse serum and 1% normal goat serum in PBS). Slides were incubated with primary antibodies overnight at 4°C and secondary antibodies for 1 hour at room temperature. For immunohistochemistry staining, the secondary antibody was biotin conjugated, the sections were treated with ABC (avidin-biotin complex) reagent (Vector Labs), and the nuclei were counterstained with hematoxylin (Sigma-Aldrich). Tumor sections were mounted with Cytoseal mounting medium (Life Technologies). Quantifications of immunohistochemistry images were assessed by examining three randomly selected low-power fields per slide. For immunofluorescence staining, tumor sections were mounted in an antifade fluorescence mounting medium with 4′,6-diamidino-2-phenylindole. Slides were visualized under a Nikon Eclipse Ti fluorescence microscope.
Cytoseal mounting medium
Manufactured by Thermo Fisher Scientific
Sourced in United States
Cytoseal mounting medium is a solvent-based media designed for use in microscopy applications. It is formulated to mount and preserve specimens on microscope slides.
Automatically generated - may contain errors
Lab products found in correlation
Hematoxylin, by Merck Group (5 mentions)
Eclipse ti, by Nikon (3 mentions)
Abc reagent, by Vector Laboratories (2 mentions)
Gill 2 hematoxylin, by Thermo Fisher Scientific (2 mentions)
Eosin y, by Thermo Fisher Scientific (2 mentions)
Hematoxylin, by Thermo Fisher Scientific (2 mentions)
Nanozoomer slide scanning system, by Hamamatsu Photonics (2 mentions)
Npr 0008615, by Advanced Cell Diagnostics (2 mentions)
Npr 0008187, by Advanced Cell Diagnostics (2 mentions)
Zikv rna probe, by Advanced Cell Diagnostics (2 mentions)
Abc avidin biotin complex reagent, by Vector Laboratories (1 mentions)
Abc reagent, by Merck Group (1 mentions)
Hybez oven, by Advanced Cell Diagnostics (1 mentions)
Basescope, by Advanced Cell Diagnostics (1 mentions)
Mouse stem cell factor, by Thermo Fisher Scientific (1 mentions)
Mouse thrombopoietin, by Thermo Fisher Scientific (1 mentions)
Mouse interleukin 3, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
22 protocols using cytoseal mounting medium
1
Immunohistochemistry and Immunofluorescence Staining of Frozen and Paraffin-Embedded Tumor Sections
2
Immunohistochemical Tissue Analysis
3
Immunohistochemical and Immunofluorescence Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen tumor sections were sequentially fixed with cold acetone, acetone plus chloroform (1:1), and acetone. Paraffin-embedded sections were deparaffinized and heated in antigen retrieval buffer. Tissue sections were blocked with 3% H2O2 in distilled water for 20 min and then in blocking buffer (5% normal horse serum and 1% normal goat serum in PBS). Slides were incubated with primary antibodies (see online supplemental file 1 ) overnight at 4°C and secondary antibodies (see online supplemental file 1 ) for 1 hour at room temperature. For immunohistochemistry staining, the secondary antibody was biotin conjugated, the sections were treated with ABC reagent (Vector Labs), and the nuclei were counterstained with hematoxylin (Sigma-Aldrich). Tumor sections were mounted with Cytoseal mounting medium (Life Technologies). Quantifications of immunohistochemistry images were assessed by examining three randomly selected low-power fields per slide. For immunofluorescence staining, tumor sections were mounted in an antifade fluorescence mounting medium with 4′,6-diamidino-2-phenylindole. Slides were visualized under a Nikon Eclipse Ti fluorescence microscope.
4
Immunohistochemical Analysis of Tumor Sections
5
Immunohistochemical and Immunofluorescence Staining of Tumor Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Frozen tumor sections were sequentially fixed with cold acetone, acetone plus chloroform (1:1), and acetone. Paraffin-embedded sections were deparaffinized and heated in antigen retrieval buffer. Tissue sections were blocked with 3% hydrogen peroxide in distilled water for 20 min and then in blocking buffer (5% normal horse serum and 1% normal goat serum in PBS). Slides were incubated with primary antibody overnight at 4°C and secondary antibody for 1 hour at room temperature. For immunohistochemical staining, the secondary antibody was biotin-conjugated, the sections were treated with avidin–biotin complex (ABC) reagent, and the nuclei were counterstained with hematoxylin (Sigma-Aldrich, St. Louis, MO). Tumor sections were mounted with Cytoseal mounting medium (Life Technologies, Carlsbad, CA). Immunohistochemical staining was quantified in three randomly selected low-power fields (20×) per slide. For immunofluorescence staining, tumor sections were mounted in antifade with 4′,6-diamidino-2-phenylindole fluorescence mounting medium. Slides were visualized under a Nikon Eclipse Ti fluorescence microscope.
6
Histological Analysis of Aortic and Adipose Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The paraffin-embedded aortic and adipose tissues were deparaffinized twice in xylene and rehydrated in ethanol before being sectioned (Leica, Leider Lane, Wetzlar, Hesse, Germany) at 5 µm thick. Thereafter, the aortic and adipose tissue sections were stained with hematoxylin and eosin (H&E) (Merck, Frankfurter, Darmstadt, Germany). All slides were mounted with Cytoseal mounting medium (Richard-Allan Scientific™, Kalamazo, MI, USA). The images were obtained by light microscope (Olympus BX41, Shinjuku-ku, Tokyo, Japan) at 1000× magnification for the aortic section to visualize the endothelial surface and alignment of aortic elastic lamina, and at 400× magnification for the adipose tissue section to assess the adipocyte size. Image J Software (ImageJ, NIH-Bethesda, Maryland, USA) was used to analyze the surface area, perimeter, and number of adipocyte in the adipose tissue.
7
HER2 Expression Analysis by RNAscope
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Viable cells were recovered from the filters by backwashing as described above, pelleted, then washed in PBMC wash (Advanced Cell Diagnostics), and subsequently suspended in 70% ethanol. Cells collected from one filter were cytospun onto Octospot slides (Thermal Fisher) using a Shannon Cystospin Centrifuge, such that cells from each filter was divided into 8 spots. The slides were then treated sequentially with a peroxidase and protease according to the manufacturer’s recommendations. RNAscope 2.0 HD assays were performed using target probe Hs-HER2 (Advanced Cell Diagnostics). The assays were performed manually according to the manufacturer's instructions. The target probes were hybridized for 2 hours in a hybridization oven at 40°C before performing amplification steps according to the manufacturer. Signals were generated by chromogenic reaction using horseradish peroxidase with 3, 3-diaminobenzidine. The slides were counterstained with hematoxylin and mounted with Cytoseal mounting medium (Richard-Allan Scientific). Positively stained cells were counted manually.
8
Paraffin Sectioning and Hematoxylin-Eosin Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fixed pancreatic tissue was embedded in paraffin and sectioned into 4 μm-thick sections. Tissue sections were then stained with hematoxylin and eosin. Sections were deparaffinized with xylene and rehydrated in decreasing concentrations of ethanol. Sections were stained with Gill 2 Hematoxylin (Richard-Allan Scientific, San Diego, CA, USA, 72504) and Eosin-Y (Richard-Allan Scientific, 71204) according to manufacturer’s protocols. Sections were dehydrated in increasing concentrations of ethanol, cleared in xylene, dried and coverslipped with Cytoseal Mounting Medium (Richard-Allan Scientific, 48212–187).
9
BaseScope Assay for Splice Variant Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The BaseScope™ (Advanced Cell Diagnostics, CA, USA) assays were performed manually according to the manufacturer’s instructions. This method allows the detection of exon junctions and the analysis of splice variants. Briefly, the BaseScope™ assay procedure included the following steps: FFPE sections were deparaffinised and treated sequentially with specific pre-treatments to allow for target probe access. Target probes were added onto the slides and incubated in the HybEZ oven (Advanced Cell Diagnostics) for 2 h at 40 °C to allow probe hybridization to RNA targets. The slides were washed and incubated with a series of signal amplification solutions. The signal was amplified using a multi-step process, and detected using a red chromogenic substrate (10 min at room temperature). The slides were counterstained with haematoxylin and mounted with Cytoseal mounting medium (Richard-Allan Scientific, CA, USA).
10
Hematoxylin and Eosin Staining of Paraffin-Embedded Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fixed pancreatic tissue was embedded in paraffin and sectioned into 4 µm-thick sections. Tissue sections were then stained with hematoxylin and eosin. Sections were deparaffinized with xylene and rehydrated in decreasing concentrations of ethanol. Sections were stained with Gill 2 Hematoxylin (Richard-Allan Scientific, San Diego, CA, USA, 72504) and Eosin-Y (Richard-Allan Scientific, 71204) according to manufacturer’s protocols. Sections were dehydrated in increasing concentrations of ethanol, cleared in xylene, dried and coverslipped with Cytoseal Mounting Medium (Richard-Allan Scientific, 48212-187).
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!