Understanding how cells respond to external stimuli is crucial. However, there are a lack of inspection systems capable of simultaneously stimulating and imaging cells, especially in their natural states. This study presents a novel microfluidic stimulation and observation system equipped with flat-fielding quantitative phase contrast microscopy (FF-QPCM). This system allowed us to track the behavior of organelles in live cells experiencing controlled microfluidic stimulation. Using this innovative imaging platform, we successfully quantified the cellular response to shear stress including directional cellular shrinkage and mitochondrial distribution change in a label-free manner. Additionally, we detected and characterized the cellular response, particularly mitochondrial behavior, under varying fluidic conditions such as temperature and drug induction time. The proposed imaging platform is highly suitable for various microfluidic applications at the organelle level. We advocate that this platform will significantly facilitate life science research in microfluidic environments.
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