The specimens were fixed using 10% formalin (Solarbio, China) and embedded in the paraffin wax (Solarbio, China) followed by slicing. After deparaffinization using xylene (Solarbio, China) and rehydration by ethanol (Solarbio, China). The primary antibody against Ki-67 (Beyotime, China) was applied and incubated at 4°C for 16 h. The DAB Histostaining kit (Solarbio, China) was selected to localize Ki-67 in clinical tissues.
Formalin
Manufactured by Solarbio
Sourced in China
Formalin is a solution of formaldehyde in water, commonly used as a fixative agent in histological and biological sample preparation. It preserves tissue structure and cellular morphology, enabling effective analysis and examination of specimens.
Automatically generated - may contain errors
Lab products found in correlation
Ripa buffer, by Thermo Fisher Scientific (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Paraffin wax, by Solarbio (1 mentions)
Xylene, by Solarbio (1 mentions)
Ethanol, by Solarbio (1 mentions)
Dimethylbenzene, by Maclin Biochemical Technology (1 mentions)
Hydrogen peroxide, by Maclin Biochemical Technology (1 mentions)
Inverted light microscope, by Leica (1 mentions)
Bx 145 microscope, by Olympus (1 mentions)
Quant it picogreen kit, by Thermo Fisher Scientific (1 mentions)
Bx53f inverted microscope, by Olympus (1 mentions)
Mayer s hematoxylin, by Sangon (1 mentions)
Oil red o solution, by Sangon (1 mentions)
11 protocols using formalin
1
Immunohistochemical Analysis of Ki-67
2
Quantifying Hepatic Lipid and Glycogen
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Mouse livers were firstly processed in the formalin (Solarbio) and then embedded in the paraffin tissue blocks. 4 μm-thick sections were cut using a microtome, and transferred onto Bond-RiteTM slides. After being heated at 60℃ for 1 hour, the sections were deparaffinized in fresh xylene (three changes), and then rehydrated with descending grades of alcohol. Next, the sections were performed with either PAS (Solarbio) or H&E (Servicebio) staining, and comparatively viewed under the Olympus microscope equipped with an Olympus digital camera. Three areas having the same size were picked casually from each liver section, and their staining density was counted with the Image J software. The average value of lipid droplets within H&E staining or glycogen-positive area in PAS staining was utilized to respectively determine total lipid or glycogen content of mouse liver.
3
Modeling Persistent Foramen Ovale in Mice
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Breeding pairs of 129T2/SvEms mice with a PFO incidence of approximately 75% were purchased from the Jackson Laboratory.89 Mice were housed and bred in the animal SPF facility of Sichuan University. All animals were group‐housed (4 per cage) with food and water under a 12 h dark–light cycle (8:00 p.m. to 8:00 a.m.) in a temperature‐controlled room. Mice were anesthetized with 2−3% isoflurane inhalation. Retro‐orbital blood was collected from 6‐ to 8‐week‐old offspring after removing eyeballs, and the plasma was separated by low‐speed centrifugation (3000×g × 15 min). After rapid decapitation of the mice, the brains were rapidly removed, and the cerebral tissues were cut into three equally spaced (approximately 3 mm) coronal blocks stored at −80°C. Whole hearts were dissected, fixed in formalin (Solarbio) for 48 h, and embedded in wax. Next, 3.5 μm continuous coronal sections of the heart were stained with hematoxylin and eosin (H&E) to assess the presence of PFO according to a previously described procedure.90 The animal experiments were approved by the Institutional Animal Care and Use Committee of Sichuan University (20211503A) and conformed to the Guide for the Care and Use of Laboratory Animals.
4
Placental Biomarker Profiling via ELISA
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Fresh placental tissues were collected after delivery and fixed in 10% formalin (Solarbio Technology Co., Ltd., Beijing. China), and then finely ground with RIPA buffer (Thermo). Tissue residues were discarded after centrifuging. Tissue suspension was used for detecting placental indexes by enzyme linked immunosorbent assay (ELISA), including nuclear factor-κB (NF-κB), TNF-α, IL-1β, IL-6, IL-8, IL-10, leptin, adiponectin, visfatin, retinol-binding protein-4 (RBP-4), chemerin, nesfatin-1, insulin receptor substrate-1 (IRS-1), IRS-2, fatty acid transport protein-4 (FATP-4), endothelial lipase (EL), low density lipoprotein (LPL), fatty acid binding protein-1 (FABP-1), FABP-3, FABP-4 and FABP-5. ELISA kits were obtained from TaKaRa Biotechnology Co., Ltd. (Dalian, China), Beijing Wantai Biological Pharmacy Enterprise Co., Ltd. (China) and Shanghai Kehua Bio-Engineering Co., Ltd. (China).
5
Immunohistochemical Analysis of KIF21B
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Human tissues were fixed in 10% formalin (Solarbio, Beijing, China) and embedded in paraffin. Five-micrometer sections were dewaxed with dimethylbenzene (Macklin, Shanghai, China) and dehydrated in a gradient ethanol series, followed by incubation with citrate buffer for antigen recovery. Hydrogen peroxide (Macklin, 3%) was used to block the endogenous peroxidase activity. The sections were incubated with primary antibodies against KIF21B (1:500; Thermo Fisher) overnight at 4°C. The sections were then stained using biotinylated secondary antibodies at 37°C for 20 min and exposed using DAB. The images were captured by microscopy (Leica, Wetzlar, Hesse-Darmstadt, Germany). Three fields were randomly selected, and the average percentage of brown or dark yellow particles in the cytoplasm was counted.
6
Quantifying Cardiac Fibrosis and Cell Morphology
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The heart tissue samples were fixed with 10 % formalin (Solarbio Technology Co., Ltd., Beijing, China) at RT for 24 h. Subsequently, the samples were embedded in paraffin wax and sliced into 3‐μm‐thick sections. To assess the cardiac architecture and fibrosis, the H&E or Masson's trichrome stained slices were examined under an inverted light microscope (Leica, Berlin, Germany), respectively. The degree of cardiac fibrosis was quantified using ImageJ software and expressed as a percentage of the fibrotic area of the whole region. The cross-sectional area of cardiac cells was quantified using HALO software v3.5.3577.
7
Endometriosis Tissue and Blood Analysis
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Peripheral blood samples were obtained 1 day before surgery, and red fresh endometriotic lesions and endometrial tissues were collected during the surgical procedure. The blood or cell supernatants were centrifuged at 1,000 g for 10 minutes, and the supernatants were transferred into 1.5 mL tubes and stored at −80 °C until processing. Of the 37 women with endometriosis, 16 (43.2%) had ovarian endometriosis, 11 (29.7%) had peritoneal endometriosis and 10 (27.0%) had deeply infiltrating endometriosis. Half of the endometrial tissue was frozen in −80 °C for mRNA detection, and the other half was immersed in formalin (Solarbio) for further immunohistochemical and immunofluorescence staining.
8
Colony Formation Assay for OCM1 and C918 Cells
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A total of 800 OCM1 or C918 cells were cultured in 6-well plates for 14 days. Then, colonies were fixed with formalin (Solarbio, Beijing, China). The colonies containing 50 cells were counted for further analysis.
9
Histological Analysis of Liver Tissue
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For routine histology, the liver tissues were preserved in 10% formalin (Solarbio, Beijing, China) and embedded in paraffin and cut into 50-μm thick sections, which were prepared and stained with hematoxylin and eosin (H&E) and Masson staining by using standard methods. All images of pathological changes were captured with the Olympus BX-145 microscope (Olympus, Japan).
10
Alkaline Phosphatase Activity Assay
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ALP activity was detected using kits (MSK Biotechnology, Wuhan, Hubei, China). The cells were fixed with 10% formalin (Solarbio) for 20 min, fixed with the same amount of ethanol and acetone ice solution for 1 min, and washed with PBS. After that, the cells were placed in ALP solution, incubated at 37 °C for 20 min, and observed under a microscope. The cells were seeded into the 96-well plates and the optical density value was measured at 405 nm using a microplate reader (Thermo Fisher Scientific). ALP activity was then normalized to DNA content using Quant-iT PicoGreen kit (Invitrogen).
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