The experimental procedure for histological observation was referred to the protocol of our laboratory [38 ]. Samples were fixed in 4% paraformaldehyde (PFA) (Boster Biological Technology, USA) overnight, then transferred into methanol (30%, 50%, 70%, 90%, 2 h, respectively), and stored in 100% methanol. After dehydrating in ethanol and embedding in paraffin, the tissue blocks were cut into 5 μm thickness. The haematoxylin-eosin staining experiment was performed according to manufacturer’s specification (Solarbio, Beijing, China). The results were observed and photographed using an Olympus BX43 microscope (Tokyo, Japan).
Paraformaldehyde pfa
Manufactured by Boster Bio
Sourced in United States
Paraformaldehyde (PFA) is a white, crystalline solid that is commonly used as a fixative in various laboratory applications. It serves as a source of formaldehyde, which is essential for preserving and stabilizing biological samples, such as cells and tissues, for analysis and visualization.
Automatically generated - may contain errors
Lab products found in correlation
Bx43 microscope, by Olympus (1 mentions)
Apomorphine, by Absin (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Bca protein assay kit, by Boster Bio (1 mentions)
Tnf α elisa kit, by Cloud-Clone (1 mentions)
6 hydroxydopamine hydrobromide 6 ohda, by Merck Group (1 mentions)
Semaglutide, by Bachem (1 mentions)
Crystal violet, by Solarbio (1 mentions)
Dmi6000b, by Leica (1 mentions)
4 protocols using paraformaldehyde pfa
1
Histological Tissue Preparation and Staining
2
Parkinson's Disease Modeling and Analysis
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Reagents used were 6-hydroxydopamine hydrobromide (6-OHDA, Sigma, USA), pargyline hydrochloride (MedChemExpress, USA), desipramine hydrochloride (MedChemExpress, USA), apomorphine (Absin Bioscience Inc, China), paraformaldehyde (PFA, Boster Biotechnology, China), RIPA lysis buffer (Beyotime Institute of Biotechnology, China), BCA Protein Assay Kit (Boster Biotechnology, China), ECL-enhanced chemoluminescence (Boster Biotechnology, China), Dopamine ELISA kit (Cloud-clone crop, China), TNF-α ELISA kit (Cloud-clone crop, China).
Semaglutide had been purchased from Bachem (Switzerland), and DA5-CH was synthetized by China peptides. The purity of each peptide was analyzed by reversed-phase HPLC and characterized using matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry, with a purity >97%. Peptides were reconstituted in ultrapure water (Milli-Q) to a concentration of 1 mg/ml, and aliquots were prepared and stored at −20°C.
Semaglutide had been purchased from Bachem (Switzerland), and DA5-CH was synthetized by China peptides. The purity of each peptide was analyzed by reversed-phase HPLC and characterized using matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry, with a purity >97%. Peptides were reconstituted in ultrapure water (Milli-Q) to a concentration of 1 mg/ml, and aliquots were prepared and stored at −20°C.
3
Plaque Reduction Assay for HSV-1
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The plaque reduction assay was conducted following the methods reported in the literature (Li et al., 2019 (link)). Briefly, Vero cell monolayers placed in 12-well plates were infected with HSV-1 (MOI = 1.5) at 37°C. Two hours later, the viral inoculum was removed, and 1 mL of methylcellulose overlay (2% methylcellulose, with 2 × final testing concentrations of SRI in a 1:1 ratio) was added to each well and left for incubation at 37°C and 5% CO2 for 72 h. The cells were eventually fixed in 4% paraformaldehyde (PFA; Boster Biological Technology, California, USA) and stained with 1% crystal violet (Solarbio, Beijing, China) for plaque counting.
4
Scalpel Loading/Dye Transfer Assay
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The scalpel loading/dye transfer (SL/DT) assay relies on the introduction of small (MW < 900) nonpermeable dyes of lucifer yellow CH dilithium salt (LY; Sigma-Aldrich, St. Louis, MO, USA) into living cells, which are traced in their intercellular movement through the gap junctions. LY is negatively charged and it has a high fluorescence efficiency [41 (link)]. Leydig TM3 cells were seeded into 6-well plates at a density of 1.25 × 105 cells/well for 24 h. After adhesion, cells were treated with experimental doses of Lepidium sativum L., starting from 62.5 to 2000 µg/mL for 48 h. After the respective treatments, a gap junction permeable tracer LY (final concentrations: 1mg/mL in calcium- and magnesium-supplemented PBS (CaMg-PBS; pH 7.2) was added to the cells and introduced into them with three parallel cuts made by a scalpel. After 6 min of incubation in the dark, the cells were washed with CaMg-PBS and fixed with 4% paraformaldehyde (PFA; Boster, Pleasanton, CA, USA). Finally, the images were captured using the Leica Application Suite X (LAS X) software, which is suitable for the fluorescent microscope DMI 6000 B (Leica Microsystems, Wetzlar, Germany), and a DCF 345FX camera. The distance at which the LY diffuses through the gap junction of TM3 cells was evaluated using ImageJ software 1.51 (National Institute of Health, LOCI, University of Wisconsin, USA) [42 (link)].
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