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DUO90806 Sigma-Aldrich

Duolink® ImageTool



application(s)   proximity ligation assay: suitable
storage temp.   20-25°C



Experiments conducted using Duolink In Situ reagents can detect and visualize protein interactions, protein expression levels and post translational modifications at the single molecule level in fixed cells and tissue samples.
To perform a complete Duolink In Situ experiment you will need two primary antibodies (IHC or ICC/IF validated) that recognize two target epitopes. Additional reagents required include a pair of PLA® probes, one PLUS and one MINUS, your choice of Detection Reagents. Recommended reagents include Wash Buffers and Mounting Medium.
Analysis is carried out using standard immunofluorescence assay equipment. HRP/Novared is also available for bright field detection. Quick and reliable quantification can be performed using Duolink ImageTool.
Duolink ImageTool is a dedicated and user-friendly software, specifically designed for objective quantification/counting of PLA signals in images generated from fluorescence microscopy. Both cells and tissue images may be analyzed. The nuclei are automatically detected and cytoplasm size estimated, enabling single cell statistical analysis of expression levels in tissue or cell populations. Furthermore regions of interest can be defined, a feature of particular relevance when studying tissue samples. Raw imaging data can be imported directly from the four major microscope vendors (Olympus, Leica, Nikon and Zeiss) as well as tiff and jpg. The results data can easily be exported to Microsoft Excel for further evaluation.

Demo version
When running the demo version you can import your own images and use all features of the software except getting the report of the analysis. To run the software in full mode you need a USB flash drive that works as a software protection key.

Download Demo Version
Duolink ImageTool Tutorial
Duolink ImageTool User Manual
Duolink ImageTool Test Images

Find answers to commonly asked question on our Duolink FAQ page

Legal Information

Duolink is a registered trademark of Sigma-Aldrich Co. LLC

PLA is a registered trademark of Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.
Protocols & Articles


Detect, Visualize and Quantify Single Post-Translational Modifications

Duolink® In Situ products enable detection, visualization, and quantification of protein events in tissue and cell samples prepared for microscopy. The in situ PLA® technology, on which these product...
Caleb Hopkins, Product Manager, Sigma® Life Science
Biofiles Vol. 8, No. 11
Keywords: Acetylations, Amplification, Apoptosis, Cancer, Cellular processes, Clinical, Diagnostic, Diseases, Enzyme activity, Gene expression, Glycosylations, Immunohistochemistry, Immunoprecipitation, Microscopy, Nucleic acid hybridization, Peptide synthesis, Phosphorylations, Post translational modifications, Transduction, Ubiquitinations, Western blot


Create Your Own PLA® Probes

Duolink® kits use in situ PLA®, a proximity ligation assay technology, to accurately and objectively quantify individual proteins, and their interactions and modifications in unmodified cells and tis...
Keywords: Bacterial conjugations, Buffers, Peptide synthesis, Phosphorylations

Duolink® In Situ Short Instructions - Fluorescence

Note: Use open droplet reactions without a cover slip and perform all incubations in a humidity chamber. Use volumes corresponding to your reaction area, see the Reaction Volume Guide at sigma.com/du...
Keywords: Amplification, Confocal microscopy, Peptide synthesis, Phase transitions

Duolink® PLA Product Selection Guide

Getting started with Duolink® PLA is simple. The protocols are very similar to that of IF, IHC, and flow cytometry, however Duolink® PLA offers distinct advantages over these and other traditional me...
Keywords: Amplification, Bacterial conjugations, Buffers, Flow cytometry, Fluorescent microscopy, Gene expression, Immunohistochemistry, Microscopy, Peptide synthesis

Peer-Reviewed Papers


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