Cardiac tissues were fixed in 10% buffered formalin for 30 min, dehydrated overnight in 75% ethanol, and embedded in paraffin. Serial sections (4 μm) were cut for morphometric analysis of the atherosclerotic lesions. The sections were stained with H&E, Masson’s trichrome, and Periodic Acid-Schiff (PAS) for histological analysis. For immunohistochemical staining, the heart sections were deparaffinized and rehydrated. Next, the sections were blocked with 3% H2O2 in methanol for 15 min to inactivate the endogenous peroxidases and incubated overnight at 4 ℃ with the primary antibodies: TGF-β (rabbit anti-TGF-β antibody, 1:300; Proteintech, Wuhan, China), collagen I (rabbit anti-collagen I antibody, 1:1000; Proteintech), collagen III (rabbit anti-collagen III antibody, 1:1000; Proteintech), MMP2 (rabbit anti- MMP2 antibody, 1:200; Proteintech), MMP9 (rabbit anti- MMP9 antibody, 1:300; Proteintech), LOX-1 (rabbit anti-LOX-1 antibody, 1:300; Abcam, England, CD36 (rabbit anti-CD36 antibody, 1:500; Proteintech). The sections were then incubated for 30 min at room temperature with a goat anti-rabbit HRP secondary antibody (anti-rabbit Universal Immunohistochemical Detection Kit; Proteintech). All sections were examined with an Olympus B×40 upright light microscope (Olympus, Tokyo, Japan).
B 40 upright light microscope
Manufactured by Olympus
Sourced in Japan
The Olympus B×40 upright light microscope is a high-quality optical instrument designed for general laboratory use. It features a sturdy, ergonomic design and provides clear, detailed images through its advanced optical system.
Automatically generated - may contain errors
Lab products found in correlation
Rabbit anti lox 1 antibody, by Abcam (3 mentions)
Histone simple stain kit, by Nichirei Biosciences (3 mentions)
Anti rabbit universal immunohistochemical detection kit, by Proteintech (2 mentions)
Rabbit anti nrf2, by Proteintech (2 mentions)
Rabbit anti nox4, by Proteintech (2 mentions)
Rabbit anti sod, by Proteintech (2 mentions)
Rabbit anti ho 1, by Abcam (2 mentions)
Rabbit anti tgf β antibody, by Proteintech (1 mentions)
Rabbit anti collagen 1 antibody, by Proteintech (1 mentions)
Collagen 3, by Proteintech (1 mentions)
Rabbit anti collagen 3 antibody, by Proteintech (1 mentions)
Lox 1, by Abcam (1 mentions)
Rabbit anti cd36 antibody, by Proteintech (1 mentions)
Collagen 1, by Proteintech (1 mentions)
Tgf β, by Proteintech (1 mentions)
Goat anti rabbit hrp secondary antibody, by Proteintech (1 mentions)
Rabbit anti cd68 antibody, by Abcam (1 mentions)
Rabbit anti caspase 3 antibody, by Abcam (1 mentions)
Rabbit anti bax antibody, by Cell Signaling Technology (1 mentions)
Rabbit anti tgf β, by Proteintech (1 mentions)
14 protocols using b 40 upright light microscope
1
Histological Analysis of Atherosclerotic Lesions
2
Histological Analysis of Grafted Skin
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The grafted skin was acquired from transplanted mouse on the day 21 after operation and was fixed by in 10% buffered formalin overnight at room temperature. The skin were embedded in paraffin the next day. Then, skin tissue slices were deparaffinized in xylene (three times, 5 min each), rehydrated (100%, 90%, 80%, and 70% alcohol, 5 min each), dehydrated, and deparaffinized. Histological changes were detected by staining 5-μm-thick sections with hematoxylin-eosin (HE) staining. Images were acquired microscopically using a B 40 upright light microscope (Olympus, Tokyo, Japan).
3
Immunohistochemical Staining of CD68 in LTs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Paraffin-embedded LTs were cut into 5 μm-thick cross-sections and deparaffinized prior to staining using a standard protocol. For immunohistochemical staining, LTs were deparaffinized and rehydrated. Next, the sections were blocked with 3% H2O2 in methanol for 15 min to inactivate endogenous peroxidases and then incubated overnight at 4℃ with one of the following primary antibodies: CD68 (No. 28058-1-AP; Proteintech, Wuhan, China). The sections were then incubated for 30 min at room temperature with a goat anti-rabbit HRP secondary antibody (No. PK10006; Proteintech). All sections were examined under an Olympus B × 40 upright light microscope (Olympus, Tokyo, Japan).
4
Immunohistochemical Analysis of LOX-1 and CD68
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry was performed using the Histone Simple stain kits (Nichirei, Tokyo, Japan) according to the manufacturer’s instructions. Briefly, paraffin-embedded sections were deparaffinized with xylene and then rehydrated in a descending series of ethanol concentrations. The sections were treated for 15 min with 3% H2O2 in methanol to inactivate endogenous peroxidases and were then incubated at room temperature for 1 h with the primary antibodies against LOX-1 (rabbit anti-LOX-1 antibody, 1:200; Abcam, England) or CD68 (rabbit anti-CD68 antibody, 1:500; Abcam, England). All sections were analyzed using an Olympus B × 40 upright light microscope (Olympus, Tokyo, Japan). For each staining, totally 3 × 7 sections (7 mice) per group were analyzed and the representative images were presented. All image analyses were done by a blinded reviewer.
5
Histological Assessment of Cardiac Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mice were anesthetized with isoflurane (1.5%) at the beginning, when the hearts were fixed by perfusion with 10% buffered formalin and used isoflurane (4%) via a nozzle placed over the nose. The hearts were fixed overnight at room temperature, transferred into 70% ethanol, and then embedded in paraffin. Paraffin-embedded cardiac tissue slices were deparaffinized via immersion in xylene (three times, 5 min each) rehydrated in a descending alcohol series (100%, 90%, 80%, and 70% alcohol, 5 min each), dehydrated in an ascending series of ethanol (70%, 80%, 90%, and 100% alcohol, 5 min each), and deparaffinized via immersion in xylene (three times, 5 min each). Histological changes were detected by staining 5-μm-thick sections with HE, FITC-conjugated WGA, or Sirius Red stain. Images were acquired microscopically using a B × 40 upright light microscope (Olympus, Tokyo, Japan).
6
Immunohistochemical Detection of Apoptosis Markers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunohistochemistry was performed using the Histone Simple stain kits (Nichirei, Tokyo, Japan) according to the manufacturer's instructions. Briefly, paraffin-embedded sections were deparaffinised with xylene and then rehydrated in a descending series of ethanol concentrations. The sections were treated for 15 min with 3% H2O2 in methanol to inactivate endogenous peroxidases and were then incubated at room temperature for 1 h with the primary antibodies against caspase-3 (rabbit anti-caspase-3 antibody, 1 : 500; Abcam, England); caspase-9 (rabbit anti-caspase-9 antibody, 1 : 500; Abcam) and BAX (rabbit anti-BAX antibody, 1 : 500; Cell Signaling Technology). All sections were analysed using an Olympus B×40 upright light microscope (Olympus, Tokyo, Japan). For each staining, a total of 3 × 7 sections (7 rats) per group were analysed and the representative images were presented. All image analyses were undertaken by a blinded reviewer.
7
Histological Analysis of Kidney Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Kidney tissue was embedded in paraffin and cut into 4 μm sections and stained as outlined previously15 (link) using TNF-α-specific antibodies (1:400 dilution; Santa Cruz biotechnology) and FN (1:500 dilution; Santa Cruz Biotechnology). Sections were also stained with H&E and Masson’s Trichrome (Maiwei, Xiamen, China) using a standard protocol. All sections were analyzed using an Olympus B ×40 upright light microscope (Olympus, Tokyo, Japan).
8
Histological Analysis of Mouse Liver Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Liver tissues (LTs) were fixed by perfusion with 10% buffered formalin. Half of a mice’s LTs were fixed overnight at room temperature, transferred to 70% ethanol, and embedded in paraffin. Paraffin-embedded LTs slices were deparaffinized by immersing them in xylene (thrice, 5 min each), rehydrated in a descending alcohol series (100%, 90%, 80%, and 70% alcohol, 5 min each), dehydrated in an ascending series of ethanol (70%, 80%, 90%, and 100% alcohol, 5 min each), and deparaffinized via immersion in xylene (thrice, 5 min each). Histological changes were detected by staining 5-µm-thick LTs sections with hematoxylin and eosin (H&E) stain according to the manufacturer’s instructions (NO. g1120; Solarbio, Beijing, China). Images were acquired using a B × 40 upright light microscope (Olympus, Tokyo, Japan).
9
Histological Analysis of Rat Heart Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rat heart tissue from each group was stored in 10% formalin for 2 weeks, dehydrated in an ascending series of alcohols (75%, 85%, 90%, and 100% alcohol, 5 min each) and embedded in paraffin wax. The 4 μm-thick paraffin sections were sliced from these paraffin-embedded tissue blocks. Tissue sections were de-paraffinised via immersion in xylene (3 times, for 5 min each) and rehydrated using a descending series of alcohols (100%, 90%, 85%, and 75% alcohol, 5 min each). Biopsy samples were stained using Masson's trichrome stain to investigate any morphological and fibrotic changes in the heart. Blue staining represented collagen accumulation. The results were visualised using an Olympus B×40 upright light microscope (Olympus, Tokyo, Japan).
10
Immunohistochemical Analysis of Cardiac Oxidative Stress
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Coronal sections of heart tissues were fixed in 10% formalin, dehydrated in an ascending series of ethanol, and embedded in paraffin for histological evaluation. Sections were deparaffinized and rehydrated. Next, they were blocked with 3% H2O2 in methanol for 15 min to deactivate endogenous peroxidases and then cultivated overnight at 4°C with one of the following primary antibodies: rabbit anti-NOX4, (1:100; Proteintech), rabbit anti-NRF2 (1:200; Proteintech), rabbit anti-HO-1 (1:100; Abcam, Cambridge, UK), rabbit anti-SOD (1:200; Proteintech), rabbit anti- Icam-1 (1:100; Proteintech), rabbit anti-ET-1 (1:100; Proteintech), rabbit anti-eNOS (1:200; Proteintech), rabbit anti-TGF-β (1:200; Proteintech), rabbit anti-Smad3 (1:100; Proteintech), rabbit anti-Co Ⅰ (1:500; Proteintech) and rabbit anti-Co Ⅲ (1:1000; Proteintech). The sections were then incubated for 30 min at room temperature with a goat anti-rabbit HRP secondary antibody (Anti-rabbit Universal Immunohistochemical Detection Kit; Proteintech). All sections were examined using an Olympus B×40 upright light microscope (Olympus, Tokyo, Japan).
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!