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High throughput single cell calcium signalling fluorometry using meander microfluidics
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DOI：10.1016/j.snr.2026.100474.png)
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Abstract
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We describe a dynamic meander microfluidic fluorometry system to capture the calcium signalling response and activation kinetics of fluorescently dyed human embryonic kidney (HEK) cell suspensions. Our system utilises the advantages of established engineered non-invasive fluorescent dyes integrated with the multifunctional capabilities of microfluidics to induce calcium-signalling through controlled exposure to extracellular ATP at a defined spatial position for synchronised cell activation and reliable method for determining response time. We observe calcium-dependent fluorescent responses with single-cell resolution over an intermediate timescale under various extracellular ATP conditions and quantify the concentration dependency to predict a system half maximal effective concentration, EC50=6(1)μM and Hill coefficient, nH=1.2(3) for the underlying activation mechanism. We further extend the fluorescence analysis to quantify the underlying cell signalling statuses through a two-state model, capable of predicting the presence of stimulated cells as early as 1.2s when using the highest extracellular ATP concentration of 100μM.
Keywords:
Microfluidics
Calcium
Cell signalling
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