Name: Video: Beyin Hasarı Olan Farelerin Optik Sinirindeki Aksonal Hasarın Elektron Mikroskobik Analizi - Deney
Uploaded: 2025-03-04T15:35:01.000+00:00
Duration: 1 min 27 s
Description: 20 Görüntüleme. Beyin Hasarı Olan Farelerin Optik Sinirindeki Aksonal Hasarın Elektron Mikroskobik Analizi

electron microscopic analysis of axonal damage in the optic nerve of mice with brain injury

Electron Microscopic Analysis of Axonal Damage in the Optic Nerve of Mice with Brain Injury

Isolate the optic nerve from transgenic mice with brain injuries at defined intervals post-injury. The nerve contains neurons expressing a fluorescent protein.
Using fluorescence microscopy, visualize the injured nerve, which exhibits a progressive increase in axonal swellings and disconnections.
Fix the tissue to preserve cellular structures.
Rinse the tissue to remove excess fixative.
Treat with an osmium-ferrocyanide solution that binds to myelin sheath lipids and enhances contrast for imaging.
Rinse the tissue, then incubate in a heavy metal compound that binds to macromolecules and enhances contrast.
Dehydrate the tissue using increasing alcohol concentrations and embed in a resin.
Obtain transverse and longitudinal sections and treat them with heavy metal compounds, improving contrast.
Under an electron microscope, the longitudinal sections reveal a progressive development of axonal swellings with accumulated swollen mitochondria, vesicles, membrane blebbing, and disrupted cytoskeletal structures.
The transverse sections show a progressive disruption of myelin sheaths, indicating axonal degeneration.

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Research

JoVE Encyclopedia of Experiments

Neuroscience

Neuropathology

Trauma

20 Görüntüleme. Beyin Hasarı Olan Farelerin Optik Sinirindeki Aksonal Hasarın Elektron Mikroskobik Analizi

Video: Beyin Hasarı Olan Farelerin Optik Sinirindeki Aksonal Hasarın Elektron Mikroskobik Analizi - Deney

Preparation of Non-human Primate Brain Tissue for Pre-embedding Immunohistochemistry and Electron Microscopy

Despite all the technological advances at the light microscopy&#160;level, electron microscopy remains the only tool in neuroscience to examine and characterize ultrastructural and morphological details of neurons, such as synaptic contacts. Good preservation of brain tissue for electron microscopy can be obtained by rigorous cryo-fixation methods, but these techniques are rather costly and limit the use of immunolabeling, which is crucial to understand the connectivity of identified neuronal systems. Freeze-substitution methods have been developed to allow the combination of cryo-fixation with immunolabeling. However, the reproducibility of these methodological approaches usually relies on costly freezing devices. Moreover, achieving reliable results with this technique is very time-consuming and skill-challenging. Hence, the traditional chemically fixed brain, particularly with acrolein fixative, remains a time-efficient and low-cost method to combine electron microscopy with immunohistochemistry. Here, we provide a reliable experimental protocol using chemical acrolein fixation that leads to the preservation of primate brain tissue and is compatible with pre-embedding immunohistochemistry and transmission electron microscopic examination.

Here, we provide an easy, low-cost, and time-efficient protocol to chemically fix primate brain tissue with acrolein fixative, allowing for long-term preservation that is compatible with pre-embedding immunohistochemistry for transmission electron microscopy.

İmmünohistokimya ve elektron mikroskobu Ön gömme için İnsan Olmayan Primat Beyin Dokusu Hazırlanması

Despite all the technological advances at the light microscopy&#160;level, electron microscopy remains the only tool in neuroscience to examine and ...

JoVE Journal

Nörobilim

Induction of Diffuse Axonal Brain Injury in Rats Based on Rotational Acceleration

Traumatic brain injury (TBI) is a major cause of death and disability. Diffuse axonal injury (DAI) is the predominant mechanism of injury in a large percentage of TBI patients requiring hospitalization. DAI involves widespread axonal damage from shaking, rotation or blast injury, leading to rapid axonal stretch injury and secondary axonal changes that are associated with a long-lasting impact on functional recovery. Historically, experimental models of DAI without focal injury have been difficult to design. Here we validate a simple, reproducible and reliable rodent model of DAI that causes widespread white matter damage without skull fractures or contusions.

This protocol validates a reliable, easy-to-perform and reproducible rodent model of brain diffuse axonal injury (DAI) that induces widespread white matter damage without skull fractures or contusions.

Rotasyonel İvmeye Dayalı Sıçanlarda Diffüz Aksonal Beyin Hasarıİn İndüksiyonu

Traumatic brain injury (TBI) is a major cause of death and disability. Diffuse axonal injury (DAI) is the predominant mechanism of injury in a large ...

EAE Hastalık Şiddetini Değerlendirmek için Omuriliğin Histolojik Boyanması

Scoring Central Nervous System Inflammation, Demyelination, and Axon Injury in Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) is a common immune-based model of multiple sclerosis (MS). This disease can be induced in rodents by active immunization with protein components of the myelin sheath and Complete Freund&#39;s adjuvant (CFA) or by the transfer of myelin-specific T effector cells from rodents primed with myelin protein/CFA into na&#239;ve rodents. The severity of EAE is typically scored on a 5-point clinical scale that measures the degree of ascending paralysis, but this scale is not optimal for assessing the extent of recovery from EAE. For example, clinical scores remain high in some EAE models (e.g., myelin oligodendrocyte glycoprotein [MOG] peptide-induced model of EAE) despite the resolution of inflammation. Thus, it is important to complement clinical scoring with histological scoring of EAE, which also provides a means to study the underlying mechanisms of cellular injury in the central nervous system (CNS).
Here, a simple protocol is presented to prepare and stain spinal cord and brain sections from mice and to score inflammation, demyelination, and axonal injury in the spinal cord. The method for scoring leukocyte infiltration in the spinal cord can also be applied to score brain inflammation in EAE. A protocol for measuring soluble neurofilament light (sNF-L) in the serum of mice using a Small Molecule Assay (SIMOA) assay is also described, which provides&#160;feedback on the extent of overall CNS injury in live mice.

Yazar Spotlight: Multipl Sklerozda Obeziteyi Otoimmün İnflamasyona Bağlayan Yolun Açıklanması 

Experimental autoimmune encephalomyelitis (EAE) serves as an animal model of multiple sclerosis. This article describes an approach for scoring spinal cord inflammation, demyelination, and axonal injury in EAE. Additionally, a method to quantify soluble neurofilament light levels in the mice serum is presented, facilitating the assessment of axonal injury in live mice.

Deneysel Otoimmün Ensefalomiyelitte Merkezi Sinir Sistemi İnflamasyonu, Demiyelinizasyon ve Akson Hasarının Skorlanması

Experimental autoimmune encephalomyelitis (EAE) is a common immune-based model of multiple sclerosis (MS). This disease can be induced in rodents by ...

Electron Microscopic Analysis of Axonal Damage in the Optic Nerve of Mice with Brain Injury

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