Paraformaldehyde-fixed Brain

Paraformaldehyde-fixed brain refers to neural tissue preserved with paraformaldehyde, a formaldehyde-releasing fixative that stabilizes cellular architecture for anatomical and molecular analysis. In aqueous solution, paraformaldehyde generates formaldehyde that forms covalent cross-links between proteins, slowing autolysis and decomposition while maintaining relationships among neurons, glia, axons, and blood vessels. After fixation, brain sections can undergo cryoprotection, embedding, immunohistochemistry, and fluorescence or electron microscopy to examine morphology and protein distribution. This preparation supports neuroscience studies of neural circuits, development, pathology, and injury, although prolonged fixation can reduce tissue permeability or mask epitopes, making fixation conditions important for reliable labeling.

Paraformaldehyde-fixed Brain - Related Videos

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JoVE Journal - Biology

Laser Capture Microdissection of Paraformaldehyde-Fixed Mouse Liver Tissue for RNA Analysis

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2026

This protocol describes a method to isolate RNA from defined histological regions of paraformaldehyde-fixed, Optimal Cutting Temperature compound-embedded mouse liver tissue using laser capture microdissection, enabling targeted downstream gene expression analysis.

Rapid In Situ Hybridization using Oligonucleotide Probes on Paraformaldehyde-prefixed Brain of Rats with Serotonin Syndrome

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Cited by 2 •

2015

This protocol describes a rapid and simplified in situ hybridization method ideal forparaformaldehyde-prefixed brain, thus reducing the need for prolonged complex steps while using fresh frozen tissues. The method is validated using the identification of the serotonin 5-HT2A receptor gene htr2a in rats.

Visualizing Astrocyte Morphology Using Fluorescent Dye Iontophoresis in a Fixed Mouse Brain Slice

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2025

Source: Moye, S. L., et.al. Visualizing Astrocyte Morphology Using Lucifer Yellow Iontophoresis. J. Vis. Exp. (2019). This video demonstrates the use of fluorescent dye iontophoresis to study astrocyte morphology in fixed brain slices. It details the piercing of the astrocyte soma with an electrode to deliver an anionic, hydrophilic dye into the cytoplasm under a low-voltage current, enabling the dye to diffuse and label the soma and branches. Confocal microscopy is then used to visualize the...

Using Spectral Reflectometry to Determine Myelinated Axon Diameters in a Fixed Mouse Brain Slice

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2025

Source: Kwon, J., et al., Spectral Reflectometric Microscopy on Myelinated Axons In Situ. J. Vis. Exp. (2018) This video demonstrates the use of spectral reflectometry with a hyperspectral confocal microscope to analyze interference patterns and determine myelinated axon diameters in fixed brain tissue slices.

Simple Elimination of Background Fluorescence in Formalin-Fixed Human Brain Tissue for Immunofluorescence Microscopy

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Cited by 43 •

2017

Background autofluorescence of biological samples often complicates fluorescence-based imaging techniques, especially in aged human postmitotic tissues. This protocol describes how autofluorescence from these samples can be effectively removed using a commercially available light emitting diode light source to photobleach the sample prior to immunostaining.

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