Journal Article

OOPS: Object-Oriented Polarization Software for analysis of fluorescence polarization microscopy images

Nawara, Tomasz J., Mattheyses, Alexa L., Dean, William F., Albert, Rose M.

2024

Overview

Most essential cellular functions are performed by proteins assembled into larger complexes. Fluorescence Polarization Microscopy (FPM) is a powerful technique that goes beyond traditional imaging methods by allowing researchers to measure not only the localization of proteins within cells, but also their orientation or alignment within complexes or cellular structures. FPM can be easily integrated into standard widefield microscopes with the addition of a polarization modulator. However, the extensive image processing and analysis required to interpret the data have limited its widespread adoption. To overcome these challenges and enhance accessibility, we introduce OOPS (Object-Oriented Polarization Software), a MATLAB package for object-based analysis of FPM data. By combining flexible image segmentation and novel object-based analyses with a high-throughput FPM processing pipeline, OOPS empowers researchers to simultaneously study molecular order and orientation in individual biological structures; conduct population assessments based on morphological features, intensity statistics, and FPM measurements; and create publication-quality visualizations, all within a user-friendly graphical interface. Here, we demonstrate the power and versatility of our approach by applying OOPS to punctate and filamentous structures.

• Publisher: Public Library of Science,Public Library of Science (PLoS)
• Subject: Algorithms / Biology and Life Sciences / Cellular structure / Computational Biology - methods / Computer and Information Sciences / Computer programs / Engineering and Technology / Fluorescence / Fluorescence microscopy / Fluorescence polarization / Fluorescence Polarization - methods / Humans / Image processing / Image Processing, Computer-Assisted - methods / Image segmentation / Microscopes / Microscopy / Microscopy, Fluorescence - methods / Open source software / Polarization / Proteins / Research and Analysis Methods / Science / Software / Statistical analysis / Technology application / Visualization (Computers)