A Red-Emitting, Multidimensional Sensor for the Simultaneous Cellular Imaging of Biothiols and Phosphate Ions Herrero-Foncubierta, Pilar Paredes Martínez, José Manuel Girón González, María Dolores Salto González, Rafael Cuerva Carvajal, Juan Manuel Miguel Álvarez, Delia Orte Gutiérrez, Ángel Dual probes Fluorescent sensors Fluorescence lifetime imaging FLIM Cellular stress Photoreceptor cells The development of new fluorescent probes for cellular imaging is currently a very active field because of the large potential in understanding cell physiology, especially targeting anomalous behaviours due to disease. In particular, red-emitting dyes are keenly sought, as the light in this spectral region presents lower interferences and a deeper depth of penetration in tissues. In this work, we have synthesized a red-emitting, dual probe for the multiplexed intracellular detection of biothiols and phosphate ions. We have prepared a fluorogenic construct involving a silicon-substituted fluorescein for red emission. The fluorogenic reaction is selectively started by the presence of biothiols. In addition, the released fluorescent moiety undergoes an excited-state proton transfer reaction promoted by the presence of phosphate ions, which modulates its fluorescence lifetime, τ, with the total phosphate concentration. Therefore, in a multidimensional approach, the intracellular levels of biothiols and phosphate can be detected simultaneously using a single fluorophore and with spectral clearing of cell autofluorescence interferences. We have applied this concept to different cell lines, including photoreceptor cells, whose levels of biothiols are importantly altered by light irradiation and other oxidants. 2018-01-24T13:47:44Z 2018-01-24T13:47:44Z 2018-01-09 info:eu-repo/semantics/article Herrero-Foncubierta, P.; et al. A Red-Emitting, Multidimensional Sensor for the Simultaneous Cellular Imaging of Biothiols and Phosphate Ions. Sensors, 18(1): 161 (2018). [http://hdl.handle.net/10481/49157] 1424-8220 http://hdl.handle.net/10481/49157 10.3390/s18010161 eng http://creativecommons.org/licenses/by-nc-nd/3.0/ info:eu-repo/semantics/openAccess Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License MDPI