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Drug-Receptor Interactions

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Related Experiment Video

Updated: Jan 20, 2026

Study of Endoplasmic Reticulum and Mitochondria Interactions by In Situ Proximity Ligation Assay in Fixed Cells

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Published on: December 10, 2016

Proximity Ligation Assay Image Analysis Protocol: Addressing Receptor-Receptor Interactions.

Marc López-Cano^1,2, Víctor Fernández-Dueñas^3,4, Francisco Ciruela^5,6

¹Unitat de Farmacologia, Departament Patologia i Terapèutica Experimental, Facultat de Medicina i Ciències de la Salut, IDIBELL, Universitat de Barcelona, L'Hospitalet de Llobregat, Barcelona, Spain.

Methods in Molecular Biology (Clifton, N.J.)

|August 22, 2019

Summary

This study details image analysis for proximity ligation assays (PLA), a method for detecting protein-protein interactions. Accurate quantification of PLA fluorescence signals using ImageJ is crucial for reliable experimental results.

Keywords:

G protein-coupled receptor Image analysis ImageJ Protein-protein interaction Proximity ligation assay

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Area of Science:

Molecular Biology
Biotechnology
Microscopy Imaging

Background:

Proximity Ligation Assay (PLA) is a sensitive method for detecting protein-protein interactions.
Accurate quantification of PLA data is essential for reliable interpretation.
Confocal microscopy images are commonly used for PLA analysis.

Purpose of the Study:

To outline image analysis methods for extracting PLA data from confocal microscopy images.
To provide a step-by-step protocol for quantifying PLA fluorescence signals using ImageJ.
To emphasize the importance of accurate image analysis in PLA experiments.

Main Methods:

Utilizing ImageJ software for image analysis.
Implementing data import and image processing functions.
Applying analytical tools for quantification of PLA signals.

Main Results:

A detailed protocol for determining and quantifying PLA fluorescence signals is presented.
The study covers essential image analysis aspects including software, data import, and processing.
The method allows for accurate assessment of protein-protein interactions.

Conclusions:

Proper image analysis of PLA confocal images is critical for accurate data interpretation.
ImageJ provides a robust platform for quantifying PLA fluorescence signals.
This approach enhances the reliability of protein-protein interaction studies.