Miras Portugal, María TeresaOrtega De La O, FelipeGualix Sánchez, Francisco JavierPérez Sen, RaquelGarcía Delicado, EsmerildaGómez Villafuertes, María RosaPelegrín, Pablo2026-01-132026-01-132020Miras-Portugal, M.T., Ortega, F., Gualix, J., Perez-Sen, R., Delicado, E.G., Gomez-Villafuertes, R. (2020). Intracellular Calcium Recording After Purinoceptor Activation Using a Video-Microscopy Equipment. In: Pelegrín, P. (eds) Purinergic Signaling. Methods in Molecular Biology, vol 2041. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9717-6_23978-1-4939-9719-0978-1-4939-9716-9978-1-4939-9717-61064-374510.1007/978-1-4939-9717-6_23https://hdl.handle.net/20.500.14352/130097Calcium is one of the most important intracellular messengers, triggering a wide range of cellular responses. Changes in intracellular free calcium concentration can be measured using calcium sensitive fluorescent dyes, which are either EGTA- or BAPTA-based organic molecules that change their spectral properties in response to Ca2+ binding. One of the most common calcium indicators is the ratiometric dye Fura-2. The main advantage of using ratiometric dyes is that the ratio signal is independent of the illumination intensity, dye concentration, photobleaching, and focus changes among others, allowing for the concentration of intracellular calcium to be determined independently of these artifacts. In this protocol, we describe the use of Fura-2 to measure intracellular calcium elevations in single cultured cells after purinoceptor activation using a video-microscopy equipment. This method, usually known as calcium imaging, allows for real-time quantification of intracellular calcium dynamics and can be adapted to measure agonist mediated intracellular calcium responses due to the activation of different purinergic receptors in several cellular models using the appropriate growth conditions.engbook part1940-6029https://doi.org/10.1007/978-1-4939-9717-6_2331646499https://pubmed.ncbi.nlm.nih.gov/31646499/restricted access57761576Calcium imagingFura-2Ratiometric calcium dyePurinergic receptorIntracellular free calcium concentrationVideo microscopyCalcium responsesNeurociencias (Biológicas)Bioquímica (Biología)Biología molecular (Biología)2490 Neurociencias2403 Bioquímica2415 Biología Molecular2302.22 Farmacología Molecular