LoQANT: An ImageJ Plugin for Quantifying Nuclear Staining in Immunohistochemistry and Immunofluorescence

Area of Science:

• Cell Biology
• Biotechnology
• Image Analysis

Background:

• Nuclear protein abundance is crucial for cellular signaling, biological activity, and disease.
• Accurate quantification of nuclear staining is vital for research in cellular function, drug development, and understanding drug resistance.
• Manual scoring methods are inefficient, prone to bias, and lack interobserver reliability.

Purpose of the Study:

• To develop an automated, reliable, and efficient method for quantifying nuclear staining.
• To overcome the limitations of manual scoring in high-throughput biological research.
• To provide a tool for accurate assessment of nuclear protein localization and abundance.

Main Methods:

• Development of LoQANT (Localisation and Quantification of Antigen Nuclear sTaining), an open-source ImageJ plugin.
• Implementation of a single-cell-based approach to analyze nuclear signals.
• Validation across various sample types using immunohistochemical and fluorescent staining.

Main Results:

• LoQANT provides reliable and efficient quantification of nuclear signals, independent of cytoplasmic staining.
• The plugin enables accurate assessment of the proportion of cells with positive nuclear signals.
• It offers both semiquantitative and quantitative measurements of nuclear staining intensity.

Conclusions:

• LoQANT effectively addresses the limitations of manual scoring for nuclear staining quantification.
• This open-source tool enhances the efficiency and reliability of cellular studies involving nuclear protein analysis.
• LoQANT is a valuable resource for researchers in cell biology, drug discovery, and disease mechanism studies.