Tissue sections were prepared as described in immunohistochemistry. The cresyl violet solution (#G1430; Solarbio, Beijing, China) was used to stain brain sections at 56 °C for 60 min, and then rinsed briefly three times with deionised water. The samples were incubated in Nissl differentiation solution for a few seconds to 2 min, then quickly followed by dehydration with gradient alcohol as described above, cleaned with xylene I for 5 min and xylene II for 5 min, and mounted with neutral balsam. We counted and analysed the cells of Nissl staining from different groups.
Cresyl violet solution
Manufactured by Solarbio
Sourced in China
1% cresyl violet solution is a laboratory reagent used for staining biological samples. It is a dye solution containing 1% cresyl violet in an appropriate solvent, typically water or another suitable liquid. The primary function of this solution is to provide a staining agent for various microscopy and histological applications.
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Lab products found in correlation
5 protocols using cresyl violet solution
1
Nissl Staining of Brain Sections
2
Nissl Staining of Brain Tissue
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Nissl staining was performed on 3 pairs of 6-month-old WT and cKO mice as previously described (You et al., 2015a). Mice were euthanized via isoflurane inhalation and decapitated. Coronal paraffin sections of brain tissue were dewaxed and rehydrated through a gradient series of ethanol, stained in 0.1% cresyl violet solution (Solarbio Life Sciences, Beijing, China) (prepared with 0.3% glacial acetic acid and filtered) for 10 minutes, rinsed in distilled H2O, dehydrated in a gradient of ethanol, cleared in xylene, and covered with glass coverslips for examination under a light microscope. Slides were also digitized with a KFpro slide scanner (KFBio, Zhejiang, China) for further analysis.
3
Cresyl Violet Staining of Brain Tissues
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Paraffin-embedded tissues being sectioned and deparaffinized, were then stained with 30 μl of 1% cresyl violet solution (Solarbio, Beijing, China) in a wet incubator for 9 min. After washes with distilled water, Nissl Differentiation solution was added onto the sections for 2 min. Subsequently, 95% ethanol was added for swift differentiation until Nissl bodies were purple and other tissues were colorless. Finally, specimens were dehydrated by absolute ethyl alcohol (Tianjin Youpu Chemical Reagent Co., Ltd., Tianjin, China), transparentized by xylene (Solarbio, Shanghai, China) and sealed by neutral gum (Biosharp, Shanghai, China). Images of the brain tissues were captured using a light microscope (Leica, Solms, Germany).
4
Nissl Staining for Neuronal Quantification
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Paraffin-embedded brain tissues were sectioned, deparaffinized, and then stained with 30 μL of 1% cresyl violet solution (Solarbio, Beijing, China) in a wet incubator for 9 min. After rinsed with distilled water, Nissl differentiation solution was added onto the sections for 2 min. Whereafter, 95% ethanol was added to differentiate swiftly until Nissl bodies were purple and other tissues were colorless. Lastly, specimens were dehydrated by absolute ethyl alcohol, transparentized by xylene and sealed by neutral gum. Images of the damaged cortex was obtained using a light microscope (Leica). Only complete neurons were counted in the Nissl-stained sections. Four sections were randomly selected from each rat, and five fields were randomly selected from each section to count the number of total neurons and dark neuron with ImageJ21 (link). The percentage of neuron survival was calculated by Eq. 1 :
5
Cresyl Violet Staining of Brain Tissue
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Paraffin-embedded tissues, after being sectioned and deparaffinized, were then stained with 30 µL of 1% cresyl violet solution (Solarbio, Beijing, China) in a wet incubator for 9 minutes. After washing with distilled water, the Nissl differentiation solution was added to the sections for 2 minutes. Subsequently, 95% ethanol was added for swift differentiation until Nissl bodies were purple, and the other tissues were colorless. Finally, specimens were dehydrated by absolute ethyl alcohol, transparentized by xylene, and sealed with neutral gum. Images of the damaged cortex and hippocampal CA2 (0.3 mm2) were taken using a light microscope (Leica) at 40× and 200× magnification.
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