Orthogonal Cell Polarity Imaging by Multiparametric Fluorescence Microscopy González García, María del Carmen Peña-Ruiz, Tomás Herrero-Foncubierta, Pilar Miguel Álvarez, Delia Girón González, María Dolores Salto González, Rafael Cuerva Carvajal, Juan Manuel Navarro, Amparo García Fernández, Emilio Orte Gutiérrez, Ángel Acridone probes Bioimaging Quantum chemistry This work has been funded by grants CTQ2017-85658-R and CTQ2017-86125-P (MICIU/AEI/ERDF)), FQM-337 (Junta de Andalucía), and 1_PIUJA 2017-18 (Universidad de Jaén). We acknowledge the Universidad de Granada (Spain) microscopy central facilities (CICUGR) and computing time from CSIRC-UGR. MCGG thanks MICIU/AEI for a predoctoral fellowship. The cellular microenvironment is a complex medium due to high concentrations of proteins and an intertwined framework of cellular organelles. In particular, cellular micro-polarity controls several biological processes, since it modulates hydrophobic/hydrophilic interactions and, hence, recognition, signalling and binding events. In this work, we have developed an unprecedented methodology to construct accurate environment polarity images using multiparametric fluorescence microscopy, via a multi-linear calibration of orthogonal parameters: the fluorescence lifetime and the spectral shift of a series of solvatochromic dyes. For this approach, we have synthesized and fully characterized N-substituted 2-methoxy-9-acridone dyes as suitable bioimaging polarity probes. However, to fully comprehend the complex links between microenvironment polarity and the dyes’ properties, we have endeavoured a multidisciplinary approach, in which we have studied the photophysics of our fluorophores using spectroscopic tools and state-of-the-art computational chemistry. This profound knowledge permitted to use these dyes as intracellular polarity probes, quantitatively and robustly probing the microenvironment of different cellular compartments. Our new methodology may pave the way to further developments in accurate sensing of cellular microenvironment parameters. 2026-01-26T11:40:17Z 2026-01-26T11:40:17Z 2020-01-27 journal article González García, M. C.; Peña-Ruiz, T.; Herrero-Foncubierta, P.; [et al.]. (2020). Orthogonal cell polarity imaging by multiparametric fluorescence microscopy Sensors and Actuators B Chemical, 309, 127770 - April 2020 https://doi.org/10.1016/j.snb.2020.127770 0925-4005 1873-3077 https://hdl.handle.net/10481/110269 /10.1016/j.snb.2020.127770 eng http://creativecommons.org/licenses/by-nc-nd/4.0/ open access Attribution-NonCommercial-NoDerivatives 4.0 Internacional Elsevier