Lucifer Yellow Permeability Assay
Lucifer Yellow Permeability Assay - Related Videos
02:01
Assessing Pseudovirus Infection Using Luciferase Reporter Assay
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2026
Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021).This video demonstrates a luciferase-based assay to quantify pseudovirus infection in epithelial cells. The pseudovirus encodes the luciferase gene, which is expressed in infected cells. After washing, and cells are incubated with lysis buffer at an ultra-low temperature, and then thawed to induce lysis.
07:03
An in vivo Assay to Test Blood Vessel Permeability
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Cited by 335 •
2013
We are presenting an in vivo assay to test blood vessel permeability. This assay is based on intravenous injection of a dye and subsequent visualization of its diffusion into interstitial spaces.
06:37
An Assay for Permeability of the Zebrafish Embryonic Neuroepithelium
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Cited by 11 •
2012
We describe a live whole animal quantitative measurement for permeability of the embryonic zebrafish brain. The technique analyzes the ability to retain cerebrospinal fluid and molecules of different molecular weights within the neural tube lumen and quantifies their movement out of the ventricles. This method is useful for determining differences in epithelial permeability and maturation during development and disease.
12:55
High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay
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Cited by 45 •
2014
We present a rapid and inexpensive screening method for identifying transcriptional regulators using high-throughput robotic transfections and a homemade dual-glow luciferase assay. This protocol rapidly generates direct side-by-side functional data for thousands of genes and is easily modifiable to target any gene of interest.
09:48
Determining Intestinal Permeability Using Lucifer Yellow in an Apical-Out Enteroid Model
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Cited by 2 •
2022
The present protocol outlines a method that utilizes lucifer yellow in an apical-out enteroid model to determine intestinal permeability. This method can be used to determine paracellular permeability in enteroids that model inflammatory bowel diseases such as necrotizing enterocolitis.
FAQs
The tracer’s hydrophilic character makes passage through the cell layer especially informative about the spaces regulated by tight junctions. When those junctions become disrupted, more Lucifer Yellow can move between adjacent cells rather than remaining restricted to the originating chamber. Consequently, fluorescence on the receiving side serves as an indirect readout of paracellular leak and barrier compromise.
Lucifer Yellow permeability assay results depend on the condition of the cellular monolayer, not simply on the presence of fluorescence. A relatively intact barrier should limit transfer, whereas injury, inflammation, or treatment-related changes can increase passage. Interpreting the signal therefore requires relating fluorescence in the opposite chamber to the barrier condition being modeled.
Quantified fluorescence converts tracer transfer into a measurable comparison between barrier conditions. A stronger signal in the receiving chamber indicates greater passage, while a weaker signal is consistent with more restricted movement. This makes the readout useful for evaluating whether an experimental injury, inflammatory state, or treatment is associated with altered epithelial or endothelial permeability.
After the monolayer and two-chamber setup has been established, the important procedural measurement is fluorescence appearing on the chamber opposite the tracer source. That signal is quantified rather than judged only by visual inspection. Comparing the measured passage across barrier conditions allows investigators to determine whether permeability has increased or decreased in the model.
Lucifer Yellow permeability assay can be applied to intestinal, vascular, and blood-brain barrier models. These systems let investigators examine barrier behavior in distinct medical contexts while using the same general permeability readout. The method is particularly relevant when the research question concerns epithelial or endothelial integrity and how that integrity changes under experimental conditions.
In drug and treatment studies, the assay helps determine whether an intervention changes tissue-barrier permeability. Researchers can compare fluorescence-based passage across experimental conditions, then assess whether the intervention is associated with greater leak or more restricted movement. This is relevant to therapies whose effects may influence intestinal, vascular, or blood-brain barrier integrity.