The experimental procedure for Nissl staining and the determination of neurotransmitter content are described below. The brains of the zebrafish were stained with Nissl stain solution (Beijing SolarBio Science & Technology Co., Ltd., Beijing, China) according to the manufacturer’s instructions, and digital images were taken with a Nikon CiS microscope (Nikon). Image-Pro Plus software (version 6.0) was used to analyze the images and count the number of Nissl-positive cells in the periglomerular gray zone (PGZ) and ventrolateral nucleus of the semicircular torus (TSvl). The concentrations of 5-HT, DA, and NE in the brain tissues of zebrafish were detected using Enzyme-linked immunosorbent assay (ELISA) kits for 5-HT, DA and NE, according to the manufacturer’s instructions.
Nissl stain solution
Manufactured by Solarbio
Sourced in China
Nissl stain solution is a laboratory reagent used for the histological staining of nervous tissue. It selectively stains the Nissl substance (rough endoplasmic reticulum) in neuronal cell bodies, allowing for the visualization and identification of nerve cells.
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Lab products found in correlation
Ci s microscope, by Nikon (1 mentions)
Inverted microscope, by Leica (1 mentions)
Erastin, by Targetmol (1 mentions)
Malondialdehyde, by Solarbio (1 mentions)
Bca kit, by Beyotime (1 mentions)
Tublin, by Cell Signaling Technology (1 mentions)
Micro reduced glutathione gsh assay kit, by Solarbio (1 mentions)
Acsl4, by Santa Cruz Biotechnology (1 mentions)
Micro malondialdehyde mda assay kit, by Solarbio (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Reactive oxygen species assay kit, by Beyotime (1 mentions)
Iron assay kit, by Nanjing Jiancheng (1 mentions)
He staining kit, by Solarbio (1 mentions)
Eclipse e100, by Nikon (1 mentions)
5 protocols using nissl stain solution
1
Nissl Staining and Neurotransmitter Analysis in Zebrafish
2
Nissl Staining of Mouse Brain
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Nissl staining was conducted using the Nissl Stain Solution (Solarbio, America) according to the manufacturer’s instructions. Mice brain sections were stained with methylene blue stain for 10 min at 65°C, then differentiated by nissl differentiation solution for 3 min. The brain sections were subsequently treated in ammonium molybdate solution for 5 min followed by a quick rinse quickly in distilled water to avoid decolorizing. Images were taken using an inverted microscope (Lecia, Germany).
3
Ferroptosis Regulation in Nervous System
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The chemical and reagents used were Nissl stain solution (cresyl violet, G1430, Solarbio), malondialdehyde (MDA) assay kit; glutathione (GSH) assay kit, dimethyl sulfoxide (DMSO), BCA kits (Beyotime Biotechnology, Beijing), rabbit antibodies against antiglutathione peroxidase 4 (GPX4, ABclonal, USA, A13309), anticystine/glutamate antiporter (SLC7A11, ab175186, Abcom, USA), ACSL4 (sc-271800, Santa Cruz Biotechnology Inc., USA), Nrf2 (ABclonal, USA), GAPDH (1 : 1000, Cell Signaling Technologies, United States, #5174), tublin (1 : 1000, Cell Signaling Technologies, United States). Micro malondialdehyde (MDA) Assay Kit (BC0025, Solarbio Science & Technology Co., Ltd., Beijing.), and Micro Reduced Glutathione (GSH) Assay Kit (BC1175, Solarbio Science & Technology Co., Ltd., Beijing). Iron Assay Kit was obtained from Nanjing Jiancheng Co. (A039-2, China). Reactive Oxygen Species Assay Kit was obtained from Beyotime Biotechnology (S0033, Beijing). Fetal bovine serum (FBS) was purchased from GIBCO Inc. (Logan, Utah, USA). Erastin (T1765, TargetMol, USA) and RSL3 (HY-100218A, MedChemExpress, USA) were also used.
4
Spinal Cord Histopathological Analysis
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At 14 days after cell inoculation, rats were anesthetized with pentobarbital sodium (50 mg/kg, intraperitoneal), immediately perfused with ice-cold saline, and fixed with 4% paraformaldehyde. Then, the retrieved spinal cord tissue was embedded in paraffin and cut into 4 μm sections. After the deparaffinization and rehydration, the spinal cord sections were stained with an H&E Staining Kit or Nissl Stain Solution (Solarbio, CHN), according to the standard procedures. All stained sections were observed under a microscope. The inflammatory score and numbers of Nissl body were analyzed by ImageJ software (Dong et al., 2022 (link)).
5
Nissl Staining of Rat Spinal Cord Tissue
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Paraffin-embedded spinal cord tissue harvested from six rats in each group on the 21st day after surgery was cut into 4-μm-thick slices. After dehydration and rehydration, the slices were soaked in Nissl stain solution (Solarbio, Beijing, China, G1436) at 60°C for 0.5 hours, then differentiated with 95% ethanol. After dehydration with anhydrous ethanol, clearing with xylene, and mounting with neutral gum, an optical microscope (ECLIPSE E100, Nikon, Tokyo, Japan) was used to count the number of Nissl bodies.
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