Fixed renal tissues were embedded in paraffin and cut into 5-µm-thick sections. For hematoxylin and eosin (H&E) staining, the sections were deparaffinized in xylene at room temperature (RT) and stained with hematoxylin (Sangon Biotech Co., Ltd.) for 5 min and eosin (Beijing Solarbio Science & Technology Co., Ltd.) for 3 min, both at RT. For periodic acid-Schiff (PAS) staining, tissues were cut into 5-µm-thick sections. After deparaffinizing the sections as aforementioned, the tissues were stained with periodic acid for 10 min and counterstained with Schiff for 15 min, both at RT, using a PAS staining solution obtained from BaSO Biotech Co., Ltd.. All the sections were observed using a fluorescence microscope (Olympus Corporation; magnification, ×200 for H&E and ×400 for PAS). Inflammation was scored on a scale from 0 to 4: 0, None; 1, rare; 2, common; 3, frequent; and 4, severe (27 (link)). PAS-positive cells were quantified using Image Pro-Plus v6.0 (Media Cybernetics, Inc.).
Hematoxylin
Manufactured by Sangon
Sourced in China
Hematoxylin is a natural dye derived from the heartwood of the Logwood tree. It is a commonly used stain in histology and cytology labs for the visualization of cell nuclei. Hematoxylin stains cell nuclei a deep blue-purple color, allowing for the identification and examination of cellular structures.
Automatically generated - may contain errors
Lab products found in correlation
Hematoxylin and eosin, by Sangon (3 mentions)
Xylene, by Sangon (3 mentions)
Bodipy, by Merck Group (1 mentions)
F4 80, by Proteintech (1 mentions)
Oil red o, by Sangon (1 mentions)
3 3 diaminobenzidine (dab), by Beyotime (1 mentions)
Ab279653, by Abcam (1 mentions)
Normal horse serum, by Vector Biolabs (1 mentions)
3 3 diaminobenzidine solution, by Sangon (1 mentions)
Light microscopy, by Sangon (1 mentions)
3 3 diaminobenzidine (dab), by Gene Tech (1 mentions)
Survivin, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase conjugated goat anti mouse immunoglobulin g antibody, by Santa Cruz Biotechnology (1 mentions)
Dako real envision detection system, by Agilent Technologies (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Paraformaldehyde (pfa), by Sangon (1 mentions)
Eclipse 50i microscope, by Nikon (1 mentions)
Paraffin, by Sangon (1 mentions)
Matrigel chamber, by Corning (1 mentions)
Matrigel, by Corning (1 mentions)
25 protocols using hematoxylin
1
Histological Analysis of Renal Tissue
2
Histopathological Analysis of Liver Lipid and Fibrosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liver tissues were fixed in a 4% paraformaldehyde solution for 24 h, embedded in paraffin. hematoxylin-eosin (H&E) (hematoxylin, E607317-0500; eosin, E607321-0100, Sangon Biotech, Shanghai, China) staining was performed on paraffin-embedded tissues to visualize the pattern of lipid accumulation. Oil Red O (E607319-0010; Sangon Biotech, Shanghai, China) staining was performed on optimal cutting temperature (OCT) compound (4583, Sakura, Torrance, CA)-embedded frozen liver sections to visualize lipid droplet accumulation. Sirius red (PH1098; Scientific Phygene, Beijing, China) staining was performed to detect liver fibrosis with standard techniques. F4/80 (Proteintech) staining was performed to detect liver lobule inflammation. Moreover, to characterize the hepatocytes lipid accumulation, bodipy (Sigma) immunofluorescence of primary hepatocytes, HepG2, and AML12 cells was performed. Histological features were observed and captured with a light microscope (OLYMPUS DP80, Olympus, Tokyo, Japan).
3
Immunohistochemical Analysis of Xenograft Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Formalin-fixed paraffin-embedded sections of xenografts were deparaffinized with xylene and graded ethanol. For H&E, the sections were stained with hematoxylin and eosin (Sangon Biotech) according to the standard protocol 9 . For IHC, the sections were incubated with 0.3% H2O2 in ethanol for 30 min, followed by incubation with anti-PODNL1 overnight at 4 °C. Subsequently, the sections were visualized using 3,3'-diaminobenzidine (DAB, Beyotime Biotechnology) and co-stained with hematoxylin (Sangon Biotech). Finally, the sections were mounted with neutral gum (CWBIO) and captured by the microscope (Zeiss).
4
Immunohistochemical Analysis of Ki67 Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The excised tumor samples were embedded and fixed. Sections were microwaved in Retrieval Solution (Amyjet, Wuhan, China) at pH 6.0 or pH 9.0 for 7 min, followed by incubation in 3 % H2O2 for 10 min. Nonspecifically bound sites were then blocked with 2.5 % normal horse serum (Vector Biolabs, Burlingame, CA, USA). The sections were incubated with an antibody against Ki67 (#ab279653, Abcam, USA). After incubation with a secondary antibody (#ab125913, Abcam) for 30 min, the sections were stained with 3,3′-diaminobenzidine solution (Sangon Biotech, Shanghai, China) and hematoxylin (Sangon Biotech). Representative areas were viewed under a light microscope (Nikon 90i, Tokyo, Japan).
5
Animal Tumor Histological Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Animal tumor slides heated at 56°C for 1 hour, dewaxed the slide with xylene(Sangon Biotech, shanghai, China), 95% and 75% ethanol(Sangon Biotech, shanghai, China), and water; then stained the slide with hematoxylin(Sangon Biotech, shanghai, China) for 5min, washed the slides with water for 30s, dipped in 1% hydrochloric acid (Haoran Biological technology CO., LTD, shanghai, China) for 3 sec and washed with water for 10min, and stained the slide with 1% eosin (Sangon Biotech, shanghai, China) for 3min, then washed with water. Finally dehydrated, transparented and mounted the slides.
6
Immunohistochemical Staining of Tissue Sections
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The frozen tissue section slides were fixed in acetone for 15 minutes and subsequently washed by PBS. Endogenous peroxidases and non-specific binding were blocked by incubating with 3% hydrogen peroxide for 10 minutes and with 3% BSA, respectively. Slides were incubated with primary antibodies for 30 minutes at room temperature, followed by a 30-minute incubation with horseradish peroxidase-conjugated goat anti human IgG antibodies (Thermofisher). Antibody binding was visualized by incubating with the peroxidase substrate 3,3’-diaminobenzidine (DAB, GeneTech) for 5 minutes. After counterstaining with hematoxylin (Sangon Biotechnology), tissue sections were analyzed using light microscopy (Nikon).
7
Immunohistochemical Analysis of Tumor Xenografts
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tumor xenografts were fixed in formalin solution overnight at room temperature and embedded in paraffin blocks, which were sliced into 4-µm thick sections for immunohistochemical staining. Following deparaffinization, the sections were incubated at 4°C overnight with antibodies of Bcl-2 (cat no. 15071), Bax (cat no. 5023), caspase-3 (cat no. 9662) and survivin (cat no. 2808) (1:500; Cell Signaling Technology, Inc.). The sections were washed 3 times with PBS with 0.1% Tween-20 (PBST), and incubated for 1 h at room temperature with a horseradish peroxidase-conjugated goat anti-mouse immunoglobulin G antibody (1:200; cat no. sc-2031; Santa Cruz Biotechnology, Inc.). The slices were washed with PBST 3 times, and incubated in diaminobenzidine (Sangon Biotech Co., Ltd., Shanghai, China) for 5 min at room temperature and counterstained with hematoxylin (Sangon Biotech Co., Ltd.) for 1 min at room temperature. Images were captured using a light microscope (Olympus Corporation) at a magnification of ×20. Five sections per tumor tissue were examined.
8
Immunohistochemical Analysis of HLA-DR Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gingival tissues were fixed in 4% paraformaldehyde (Sangon Biotech Co., Ltd., Shanghai, China) for 24 h and embedded in paraffin (Sangon Biotech Co., Ltd.). The tissue was cut into 4 mm sections, blocked in 0.5% bovine serum albumin (Sigma-Aldrich, St. Louis, MO, USA) for 30 min and incubated with rabbit anti-human leukocyte antigen (HLA)-DR polyclonal antibody (cat. no. ab175085; Abcam, Cambridge, UK; dilution 1:1,000) overnight at 4°C in a humid chamber, prior to being washed three times with 0.01 M phosphate-buffered saline (PBS). The tissue samples were subsequently incubated with secondary antibody (goat anti-rabbit/mouse; Dako, Glostrup, Denmark) at 37°C for 1 h, prior to being washed three times with 0.01 M PBS. The immune complex was visualized using a Dako REAL™EnVision™ Detection system containing peroxidase/DAB, according to the manufacturer's protocol. The nuclei were counterstained with hematoxylin (Sangon Biotech Co., Ltd.), and the sections were observed under a Nikon Eclipse 50i microscope (Nikon Corporation, Tokyo, Japan).
9
Transwell Assay for Ovarian Cancer Cell Invasion
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After transfection, the invasive capacity of CaOV3 and SKOV3 cells was determined using transwell assay. Briefly, the Matrigel chamber (Corning, New York, United States) used in this study was coated with 50 μl Matrigel (1:8) in advance. CaOV3 and SKOV3 cells were suspended in serum-free medium at a concentration of 5 × 104/ml. Then, 200 μl of the suspended cells was added to the upper chamber, and 600 μl of complete medium was provided to the lower chamber. After maintaining for 48 h in a 5% CO2 and 37°C incubator, the uninvaded cells on the upper chamber were wiped off, while the successfully invaded cells on the lower chamber were fixed with 95% ethanol (Aladdin) for 20 min and stained with hematoxylin (Sangon Biotech) for 10 min. Finally, the images of the invaded cells were photographed with a microscope (Olympus).
10
Histological Analysis of Liver Lipids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh liver tissues were rapidly frozen and embedded. After fixation in 4% formaldehyde, sections were stained with Oil Red O (Sigma-Aldrich) and counterstained with hematoxylin (Sangon, China). Images were captured using a Leica microscope.
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!