In Situ Fractionation
In Situ Fractionation - Related Videos
09:20
In situ Subcellular Fractionation of Adherent and Non-adherent Mammalian Cells
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Cited by 29 •
2010
In situ subcellular fractionation of mammalian cells on microscope coverslips allows the visualisation of protein localisation.
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...
08:00
Whole-Mount In Situ Hybridization
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2023
Whole-mount in situ hybridization (WMISH) is a common technique used for visualizing the location of expressed RNAs in embryos. In this process, synthetically produced RNA probes are first complementarily bound, or "hybridized," to the transcripts of target genes. Immunohistochemistry or fluorescence is then used to detect these RNA hybrids, revealing spatial and temporal patterns of gene expression. Unlike traditional in situ hybridization techniques, which require thin tissue sections whose...
07:57
Using the GELFREE 8100 Fractionation System for Molecular Weight-Based Fractionation with Liquid Phase Recovery
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Cited by 17 •
2009
The accompanying video describes the use of the GELFREE 8100 Fractionation System, which partitions complex protein samples on the basis of molecular weight and recovers the fractions in liquid phase. The video describes how the technology works, how it is used, and provides resultant data, with polyacrylamide gel electrophoresis analysis of fractionated bovine liver homogenate.
04:16
Stromal Vascular Fraction-Enriched Fat Grafting for the Management of Symptomatic Neuromas
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2025
Source: Zimmermann, S., et.al. Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata. J. Vis. Exp. (2017)This video demonstrates the surgical procedure for neurolysis and stromal vascular fraction (SVF)-enriched fat grafting in the management of symptomatic neuroma. Neurolysis removes adhesion tissue to restore nerve gliding, while SVF-enriched fat grafting is applied around the nerve to create a protective barrier, support nerve healing, and enhance...
FAQs
Selective permeabilization opens particular cellular barriers under controlled extraction conditions, allowing soluble cytoplasmic material to be released while retaining components associated with membranes, nuclei, or chromatin. This staged access is important because it preserves spatial information during processing. The resulting fractions can therefore be compared to determine whether a biomolecule is freely soluble or more closely associated with a cellular compartment.
Sequential extraction separates material in successive steps according to differences in solubility and association. Early extracts can distinguish soluble cytoplasmic components from material requiring stronger or different extraction conditions, while later fractions may contain membrane-associated, nuclear, or chromatin-bound components. This progression provides more information than a single cell lysate because it reveals relative association with intracellular structures.
Centrifugation helps partition extracted material according to physical properties after each extraction step. It supports the separation of soluble material from cellular fractions that remain associated with membranes, nuclei, or chromatin. Interpreting the resulting distribution alongside the extraction sequence allows researchers to assess where a protein or other biomolecule resides and how tightly it is associated with a compartment.
The fraction in which a biomolecule appears provides an indirect indication of its association with cellular structures. Material recovered under milder, soluble-extraction conditions is consistent with a less persistent association, whereas material retained until later extraction steps is more closely linked to membranes, nuclei, or chromatin. Comparing these patterns across conditions can reveal changes in intracellular organization or binding.
A typical workflow preserves the cells or tissue during initial processing, applies selective permeabilization, performs sequential extractions, and uses centrifugation to separate the resulting material. Researchers then examine the distribution of proteins or other biomolecules across the fractions. Keeping the extraction order consistent is essential because each step influences which components remain available for subsequent separation.
Researchers can apply in situ fractionation when they need to compare intracellular distributions before and after an experimental or environmental treatment. Shifts among soluble cytoplasmic, membrane-associated, nuclear, or chromatin-bound fractions may indicate altered localization or compartment association. This makes the approach useful for investigating signaling responses, protein trafficking, and changes in cellular organization.
The fractionation pattern can help determine where proteins and other biomolecules reside and whether their compartmental distribution changes. In biology, these observations support studies of intracellular signaling, protein trafficking, gene regulation, and cellular responses to treatment. Preserving information about original organization also helps connect biochemical measurements with the structural context of intact cells or tissues.