Axonal Fraction Isolation
Axonal Fraction Isolation - Related Videos
07:06
Isolation and Quantification of Axonal mRNAs Using Porous Membrane Inserts and RTddPCR
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Cited by 1 •
2026
This study presents a robust method to isolate axonal mRNAs using porous membrane inserts, enabling total neuron vs. neurite separation and RNA purification. Combined with RTddPCR, the approach allows absolute quantification of low-copy transcripts, facilitating studies of mRNA transport and local translation with high sensitivity, reproducibility, and broad experimental applicability.
03:24
Two-Photon Laser Axotomy: A Method to Injure Axons in Zebrafish Embryos and Observe Axonal Recovery
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2023
This video describes the method to injure axons in zebrafish embryos using two photon laser axotomy and observing axonal recovery from injury.
10:09
Fractional Distillation
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2023
Source: Laboratory of Dr. Nicholas Leadbeater — University of Connecticut Distillation is perhaps the most common laboratory technique employed by chemists for the purification of organic liquids. Compounds in a mixture with different boiling points separate into individual components when the mixture is carefully distilled. The two main types of distillation are "simple distillation" and "fractional distillation", and both are widely used in organic chemistry laboratories. Simple distillation...
10:22
Isolation of Viral Replication Compartment-enriched Sub-nuclear Fractions from Adenovirus-infected Normal Human Cells
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Cited by 7 •
2015
We provide a novel strategy to isolate viral replication compartments (RC) from adenovirus (Ad)-infected human cells. This approach represents a cell-free system that can help to elucidate the molecular mechanisms regulating viral genome replication and expression as well as regulation of viral-host interactions established at the RC.
09:31
Measurement of Tension Release During Laser Induced Axon Lesion to Evaluate Axonal Adhesion to the Substrate at Piconewton and Millisecond Resolution
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Cited by 1 •
2013
We measured the tension release in an axon that was partially lesioned with a laser dissector by simultaneous force spectroscopy measurement performed on an optically-trapped probe adhered to the membrane of the axon. The developed experimental protocol evaluates the axon adhesion to the culture substrate.
FAQs
The separation relies on physical differences among axonal material and other cellular components. During centrifugation, particles move according to their sedimentation behavior, which reflects properties such as size and density. These differences allow axon-enriched material to separate from fractions containing other neuronal or tissue components, creating a basis for more focused biochemical analysis.
Controlled homogenization prepares neuronal tissue or cell preparations for fractionation by disrupting the starting material in a regulated manner. This step must support the release and subsequent enrichment of axonal components while maintaining a basis for separating them from other cellular fractions. Its role is therefore linked directly to the quality and interpretability of the final preparation.
Both approaches use sedimentation behavior to separate cellular material, but they organize the separation differently. Differential centrifugation separates components through successive centrifugation-based fractionation, whereas density-gradient separation uses differences in density within a gradient. Either approach can contribute to axonal enrichment, depending on how the neuronal preparation is being fractionated and analyzed.
Neuronal tissue contains multiple compartments whose molecular contents can produce overlapping experimental signals. Enriching axonal material helps investigators examine changes more specifically associated with axons rather than with cell bodies, dendrites, or surrounding tissue. This distinction is especially important when interpreting axonal composition, signaling, transport-related biology, injury responses, or degeneration.
A typical workflow begins with neuronal tissue or a cell preparation, followed by controlled homogenization. The resulting material then undergoes differential centrifugation or density-gradient separation to enrich axonal components and remove other fractions. The collected material can subsequently be examined using biochemical or molecular assays, depending on the biological question and the molecules of interest.
The enriched preparation can support analysis of several axon-associated molecular categories, including proteins, RNA, organelles, and signaling molecules. Researchers may apply biochemical or molecular assays to characterize these components. Examining multiple categories can help connect axonal composition with functional processes and reveal changes that would be difficult to assign confidently in an unfractionated neuronal sample.
Axonal fraction isolation is useful when a study focuses on biological events occurring in axons rather than across the entire neuron or tissue. Applications include examining neuronal development, axonal transport, injury-related changes, and degeneration. By concentrating analysis on axon-enriched material, the method supports comparisons of axonal molecular composition and function across these contexts.