Near infrared dyes as lifetime solvatochromic probes for micropolarity measurements of biological systems

Abstract

The polarity of biological mediums controls a host of physiological processes such as digestion, signaling, transportation, metabolism, and excretion. With the recent widespread use of near-infrared (NIR) fluorescent dyes for biological imaging of cells and living organisms, reporting medium polarity with these dyes would provide invaluable functional information in addition to conventional optical imaging parameters. Here, we report a new approach to determine polarities of macro- and microsystems for in vitro and potential in vivo applications using NIR polymethine molecular probes. Unlike the poor solvatochromic response of NIR dyes in solvents with diverse polarity, their fluorescence lifetimes are highly sensitive, increasing by a factor of up to 8 on moving from polar to nonpolar mediums. We also established a correlation between fluorescence lifetime and solvent orientation polarizability and developed a lifetime polarity index for determining the polarity of complex systems, including micelles and albumin binding sites. Because of the importance of medium polarity in molecular, cellular, and biochemical processes and the significance of reduced autofluorescence and deep tissue penetration of light in the NIR region, the findings reported herein represent an important advance toward using NIR molecular probes to measure the polarity of complex biological systems in vitro and in vivo.

FIGURE 1

Structures of NIR fluorescent probes used in this study.

FIGURE 2

Normalized emission spectra of polymethine dyes and bacteriochlorophyll at 720 nm (700 nm for DTTCI).

FIGURE 3

(A) Lippert-Magada plots of N-phenyl-N-methyl-6-aminonaphtalene-2-sulfonate (25; reproduced from that reference) and LS-277. (B) Change in lifetime versus spectral shift in emission between DMSO and water (DMSO and methanol for bacteriochlorophyll). (C) Lifetime of cyanine probes versus solvent orientation polarizability. (1) Water, (2) methanol, (3), ethanol, (4) acetone, (5) DMSO, (6) methylene chloride, and (7), chloroform.