Fluorescent probes to track complex membrane blebbing

Cellular blebbing, pivotal in processes such as apoptosis, cytokinesis, and migration, involves dynamic interactions between the actomyosin network and microtubules. However, existing probes inadequately capture these simultaneous interactions, limiting our ability to study blebbing mechanisms. We present AztecBleb probes (AztecBleb^DAPI, AztecBleb^GFP, AztecBleb^TxR, AztecBleb^Cy5), novel fluorescent reporters designed to selectively target and monitor blebbing in real-time. These probes incorporate a pregnenolone-based scaffold as a hydrophobic core derived from abiraterone acetate, facilitating precise localization to blebs and microtubules without disrupting cellular function. Through persistent staining of cell blebs, these photostable and biocompatible probes enable continuous monitoring of blebbing and microtubule dynamics during cellular migration. Our approach provides new insights into the coordination of bleb formation and cytoskeletal remodeling, offering a unique tool for studying motility-driven cellular behaviors.