TipQuant: a robust algorithm for quantitative analysis of spatiotemporally dynamic activities in tip-growing cells

Cell polarity and tip growth rely on the dynamic spatial organization of signaling and structural components. Quantitative characterization of these spatiotemporal dynamics is critical for understanding polarized cell growth, yet manual quantification is labor-intensive and existing computational tools often lack the flexibility and robustness needed to analyze molecular and structural dynamics in tip-growing cells. Tip Quantification (TipQuant) identifies the cell apex by detecting the site of maximum expansion and automatically quantifies fluorescence distribution along the plasma membrane and within the apical cytoplasm from live-cell imaging data, enabling analysis of the spatiotemporal dynamics of molecular and structural components in tip-growing cells. TipQuant accurately identified cell apices and quantified the spatiotemporal behavior of fluorescently labeled proteins and cellular structures in Arabidopsis thaliana pollen tubes and Fusarium graminearum hyphae, reproducing manual measurements while reducing user bias and improving efficiency, consistency, and analytical flexibility. The tool also revealed a strong positive correlation between rho-like GTPase from plants activity and apical Ca²⁺ influx in Arabidopsis pollen tubes, demonstrating its utility for analyzing dynamic cellular processes. TipQuant is a robust analytical tool for quantifying spatiotemporal dynamics in tip-growing cells, providing a flexible alternative to manual image analysis and enabling studies of the molecular mechanisms underlying polarized growth.