AMEBaS

AMEBaS automates quantitative analysis of cell polarity from ratiometric fluorescence time-lapse imaging to extract spatiotemporal profiles of intracellular dynamics and growth in polarized cells such as pollen tubes and root hairs.

Key Features:

• Automation: Automates midline extraction to remove manual tracing and reduce bias in analysis of polarized cells.
• Segmentation: Segments the cell from background using pixel intensity thresholding to produce a binary mask isolating the cell.
• Midline Tracing: Employs skeletonization to trace the midline along the cell's major axis, providing a precise path for assessing intracellular dynamics.
• Ratiometric Kymograph Generation: Processes two-channel ratiometric time-lapse images to generate a 1D spatial kymograph along the midline that visualizes and quantifies fluorescence intensity changes over time.
• Ratiometric Analysis: Uses two fluorescence channels to correct for uneven reporter distribution without relying on arbitrary background subtraction.

Scientific Applications:

• Cell Division and Growth: Quantifies polarity establishment and maintenance during cell division and tip growth in pollen tubes and root hairs.
• Migration Studies: Measures spatiotemporal polarity dynamics relevant to cellular migration processes.
• Disease Research: Investigates disrupted cell polarity implicated in cancer and gastric dysplasia.

Methodology:

The method is based on a model of cell polarity using pollen tube and root hair growth and cytosolic ion dynamics, and it has been benchmarked with data from genetically encoded fluorescent reporters.