(1100-A) SPAchip® technology: A novel reliable method for long-term intracellular calcium detection in real time
Monday, February 5, 2024
12:00 PM – 1:00 PM EST
Location: Exhibit Halls AB
Abstract: Free calcium ions (Ca2+) play a vital role in many physiological processes and regulate some cellular functions. Thus, cytosolic calcium homeostasis is crucial to conserve cell health and avoid the excess of intracellular calcium which can lead to programmed cell death or necrosis. Depending on cell stimuli, ion fluxes occur in short times, from milliseconds to seconds or, conversely, over sustained and longer periods of time, from either intracellular compartments or the extracellular medium . Therefore, the possibility of measuring calcium concentration with a reliable and non-toxic method in dynamic and long-term conditions is important for an accurate characterization of the effects of new drugs. Here we introduce a novel technology for reliable free calcium detection that overcomes the complexity of these measurements with the current methods. With our SPAchip® technology, calcium-sensing microchips are internalized inside living cells with the easiness of any other reagent. Acting like small laboratories, our SPAchips measure reversibly calcium concentrations in real time by fluorescence emission which can be collected through several fluorescence microscopies providing with high versatility. We validated our SPAchip® technology by detecting intracellular calcium concentration in different cell lines: HL1 cardiac muscle cells, Cal51 breast cancer cells and SHSY5Y neuroblastoma cells. Besides that, we have also successfully demonstrated internalization in zebra fish embryo, an animal model which opens potential opportunities for drug discovery. By High Content Screening analysis, cell cultures with SPAchips were traced for 24 h to 5 days without compromising cell viability. Intracellular ATP induced calcium release was also measured and different drugs were used as positive controls of the experiments such as doxorubicin and 4BrA23187 ionophore, demonstrating the reliability of the method and its big contribution to the field of single cell analysis. In summary, SPAchip® technology allows the dynamic measurement of intracellular calcium concentrations over long periods of time in living cells. The technology is compatible with other cellular information including morphological parameters and comparison with extracellular media as it remains in the cytosol without producing cytotoxic effects. With great versatility in several fluorescence reading platforms, this technology has high potential not only in vitro but also in vivo applications.