Protein condensates, formed through liquid-liquid phase separation (LLPS), are implicated in various diseases including neurodegenerative disorders, cancer, and cardiovascular diseases. Despite the significance of protein condensates, the mechanisms governing their formation and microenvironmental properties remain insufficiently understood due to the lack of effective analytical tools. In this review, we explore the detection techniques and underlying mechanisms related to protein condensates, focusing on key microenvironmental properties such as viscosity, polarity, and pH. The influence of phase separation on these microenvironmental factors and its role in protein aggregation are explored. We also discuss current methods for protein labeling and imaging, comparing their advantages and limitations, and highlight their applications in live-cell studies. This review provides a comprehensive understanding of the factors influencing protein condensate dynamics and their relevance in disease, aiming to advance the development of targeted therapeutic strategies for condensate-related disorders.
Keywords: Fluorescent probes; Liquid-liquid phase separation; Microenvironment; Protein condensates.
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