Highly Potent Fluorenone Azine-based ESIPT Active Fluorophores for Cellular Viscosity Detection and Bioimaging Applications.

Abstract

Herein, synthesizes of fluorenone azine-based Schiff fluorescence probes: (E)-2-(((9H-fluoren-9-ylidene)hydrazineylidene)methyl)-5-(diethylamino)phenol (3a), (E)-9-(((9H-fluoren-9-ylidene)hydrazineylidene) methyl)-2,3,6,7-tetrahydro-1H,5H-pyrido[3,2,1-ij]quinolin-8-ol (3b), and (E)-1-(((9H-fluoren-9-ylidene)hydrazineylidene)methyl) naphthalen-2-ol (3c). The probes were structurally characterized using Fourier-transform infrared spectroscopy (FTIR), ¹H and ¹³C nuclear magnetic resonance (NMR) spectroscopy and high-resolution mass spectrometry (HRMS) analysis. The probes exhibit hydrogen bonding between phenolic -OH and imine nitrogen, enabling excited state intramolecular proton transfer (ESIPT) and free rotation in the azine (> C = N-N = C <) functional, facilitating twisted intramolecular charge transfer (TICT), and a positive solvatochromism in solvent-dependent emission studies. Further, density functional theory (DFT) based calculations accounted for the observed photophysical TICT and ESIPT processes, revealing a non-covalent interaction between phenolic -OH and imine nitrogen. Furthermore, the fluorescence intensity (log I) showed good linearity (R² = 0.999) with the viscosity (log η) with Förster-Hoffmann coefficient (X) values of 2.238, 1.405 and 3.121 for 3a, 3b and 3c, respectively. The study established the probes toxicity and fluorescence imaging in the Caenorhabditis elegans model. Probe 3a, the first azine-based probe for micro viscosity detection, demonstrated exceptional efficacy in detecting intercellular viscosity and facilitating bioimaging applications.