A Red-Emitting, Multidimensional Sensor for the Simultaneous Cellular Imaging of Biothiols and Phosphate Ions
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AuthorHerrero-Foncubierta, Pilar; Paredes Martínez, José Manuel; Girón González, María Dolores; Salto González, Rafael; Cuerva Carvajal, Juan Manuel; Miguel Álvarez, Delia; Orte, Ángel
Dual probesFluorescent sensorsFluorescence lifetime imagingFLIMCellular stressPhotoreceptor cells
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]
SponsorshipThis work was funded by grants CTQ2014-56370-R, CTQ2014-53598, and CTQ2014-55474-C2-2-R from the Spanish Ministry of Economy and Competitiveness and the European Regional Development Fund (ERDF), and grant FQM2012-790 from the Consejería de Innovación, Ciencia y Empresa (Junta de Andalucı́a), including costs to publish in open access.
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.