Post‐fixed spinal cords (n = 3 per group) were cut into 30‐μm‐thick coronal sections with cryomicrotome and mounted immediately on gelatine‐coated glass slides. Sections were Nissl‐stained to reveal neuronal cells, using the FD Cresyl Violet Solution™ (FD NeuroTechnologies, Columbia, MD, USA), following the manual of the manufacturer. The regions of interest were localized with an atlas (http://mousespinal.brain-map.org ). The images of spinal cords were captured by microscope (DMI6000B, Leica). Nissl‐positive neurons were counted from adductor motoneurons of lamina 9 (Ad9) and quadriceps motoneurons of lamina 9 (Q9) in lumbar 2–3 region. Five consecutive sections of the spinal cord from each mouse were analyzed. The study was blinded by covering the labels of slides with random numbers until statistical analysis.
Fd cresyl violet solution
Manufactured by FD NeuroTechnologies
Sourced in United States
FD cresyl violet solution is a laboratory reagent used for histological staining. It is a purple dye that stains Nissl bodies in neurons, allowing for the visualization of cell bodies in nervous tissue samples. The solution is commonly used in neuroscience research and neuropathology for the analysis of neural structures and cellular morphology.
Automatically generated - may contain errors
Lab products found in correlation
Dmi6000b, by Leica (1 mentions)
Normal blocking serum, by Vector Laboratories (1 mentions)
Normal donkey serum (nds), by Jackson ImmunoResearch (1 mentions)
3 3 diaminobenzidine (dab), by Merck Group (1 mentions)
Permount, by Thermo Fisher Scientific (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Vectastain elite abc kit, by Vector Laboratories (1 mentions)
Rabbit anti iba1 antibody, by Fujifilm (1 mentions)
5 protocols using fd cresyl violet solution
1
Nissl Staining of Spinal Cord Neurons
2
Nissl Staining of Paraffin-Embedded Brain Tissue
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For the Nissl staining, paraffin-embedded brain tissue sections (5 μm) were immersed in xylene (5 min, 2 times), rehydrated in absolute ethanol (5 min, 2 times) followed by 95%, 75%, and 50% solutions of ethanol in water (5 min each), and then washed in distilled water for 2 times, 5 min each. Slides were stained in FD cresyl violet solution (FD Neurotechnologies, Baltimore, MD, USA) for 10 min and, then, briefly rinsed in 100% ethanol and differentiated in 100% ethanol containing 0.1% glacial acetic acid for 1 min. The slides were then dehydrated in absolute ethanol (2 min, 4 times) followed by clearance in xylene (3 min, 2 times). Coverslips were mounted with resinous mounting medium. The staining of the hippocampal CA1 region was routinely analyzed by a researcher blinded to the experimental protocol.
3
Iba1-Immunohistochemistry Protocol for Microglial Staining
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The 2nd sections were processed for Iba1-immunohistochemistry. Briefly, after endogenous peroxidase activity was blocked with 0.6 % hydrogen peroxidase, free-floating sections were incubated in 0.01 M phosphate-buffered saline (PBS, pH 7.4) containing 1% normal donkey serum (Jackson ImmunoResearch, West Grove, PA), 0.3 % Triton X-100 (Sigma, St. Louis, MO), and polyclonal rabbit anti-Iba1 antibody (1:10,000, Wako Chemicals USA, Richmond, VA #019−19741) for 65 h at 4 °C. The sections were then incubated in PBS containing Triton-X, normal blocking serum, and biotinylated secondary antibody for 1 h at room temperature, and then in PBS containing avidin-biotinylated HRP complex for another 1 h using the Vectastain Elite ABC Kit (Vector Laboratories, Burlingame, CA). Subsequently, the sections were incubated for 5 min in 0.05 M Tris buffer (pH 7.2) containing 0.03 % 3′,3′-diaminobenzidine (DAB) as a chromogen (Sigma-Aldrich, St. Louis, MO) and 0.0075 % hydrogen peroxide. All sections were rinsed in distilled water, mounted on slides and were counterstained with FD cresyl violet solution (FD NeuroTechnologies, Inc.). After dehydration in ethyl alcohol, sections were cleared in xylene and cover-slipped in Permount (Fisher Scientific).
4
Cresyl Violet Staining of Brain Tissues
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The brain tissues of SN were stained with FD cresyl violet solution (FD Neurotechnologies, Inc., MD, USA) and rinsed briefly in distilled water, then differentiated in 95% ethanol containing 0.1% glacial acetic acid for 1 min. The tissues were dehydrated in 100% ethanol for 2 min and placed in xylene, then coverslipped.
5
Histopathological Analysis of Ferret Spinal Cord
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