Co-localization Assay
Co-localization Assay - Related Videos
12:55
Assay for Phosphorylation and Microtubule Binding Along with Localization of Tau Protein in Colorectal Cancer Cells
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Cited by 11 •
2017
This manuscript describes standard protocols for measuring tau hyperphosphorylation, measuring tau binding to microtubules, and localization of intracellular tau following drug treatments. These protocols can be used repetitively for screening drugs or other compounds that target tau hyperphosphorylation or microtubule binding.
02:38
Local Heat Probe Assay: A Method to Assess Nociception of High Temperatures in Drosophila Larvae
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2023
Drosophila larvae have dedicated sensory neurons–nociceptors–whose response to potentially harmful thermal stimuli triggers aversive–or so-called "nocifensive"–behaviors in the animal. This video describes a method to assess thermal nociception by recording larval behavior in response to local application of a heat probe onto the cuticle. This method is, therefore, called the local heat probe assay.
11:12
Microinjection Wound Assay and In vivo Localization of Epidermal Wound Response Reporters in Drosophila Embryos.
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Cited by 10 •
2013
The embryonic epidermis of very late stage Drosophila embryos provides an in vivo system for rapid puncture wound response analysis and can be combined with genetic manipulations or chemical microinjection treatments to advance studies in wound healing for translation into mammalian models.
09:39
Proximity Ligand Assay to Localize Proteins in DNA Damage Sites
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2024
Here, we present a simple and rapid protocol for the detection of protein interaction at DNA damage sites.
13:13
Nano-fEM: Protein Localization Using Photo-activated Localization Microscopy and Electron Microscopy
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Cited by 14 •
2012
We describe a method to localize fluorescently tagged proteins in electron micrographs. Fluorescence is first localized using photo-activated localization microscopy on ultrathin sections. These images are then aligned to electron micrographs of the same section.
FAQs
These measures convert visual overlap into quantitative evidence. Intensity correlation evaluates whether signal strengths from different fluorescent channels vary together, while shared pixel distribution assesses how much of the detected signal occupies the same image locations. Using such measures helps compare spatial patterns systematically rather than relying only on visual inspection of microscopy images.
Distinct fluorescent probes allow multiple targets to be visualized separately within the same cellular image. Each probe produces a signal associated with a particular molecule, structure, or genetic marker, and the resulting channels can then be compared for spatial correspondence. This separation is essential for determining whether the targets occupy overlapping cellular or subcellular regions.
A mutation or changed cellular condition may alter where a gene product or other target appears within the cell. Comparing localization patterns across these conditions can reveal whether spatial distribution changes, remains similar, or shifts between cellular compartments. Such differences provide evidence that the tested genetic or cellular change affects localization, without independently establishing a direct molecular interaction.
Spatial overlap shows that two signals occupy the same observed region, but it does not establish physical contact between the corresponding molecules. Targets may share an organelle, molecular complex, or broader cellular compartment without binding one another. For this reason, co-localization findings are commonly interpreted together with complementary biochemical or functional tests.
A typical workflow labels the targets of interest with distinct fluorescent probes, acquires fluorescence microscopy images, and compares the resulting signal distributions. Investigators then examine overlap between channels and may calculate intensity correlation or shared pixel distribution. The analysis can be applied to molecules, cellular structures, or genetic markers in defined cellular or subcellular regions.
In genetics, the assay helps map gene products within cells and determine whether their signals converge in particular organelles or molecular complexes. Researchers can also compare localization after mutations or altered cellular conditions. These applications connect genetic changes with spatial behavior, providing cellular context for interpreting how gene products are distributed and where they may function.
The assay can indicate whether two or more targets occupy overlapping cellular or subcellular locations and whether that pattern changes across experimental conditions. It can therefore support conclusions about shared localization, compartmental distribution, or altered positioning of gene products. The findings describe spatial relationships and should not be presented alone as proof of direct molecular association.