Lifetime‐Based Responsive Probes: Design and Applications in Biological Analysis

Abstract

With the development of modern biomedicine, biological analysis and detection are very important in disease diagnosis, detection of curative effect, prognosis and prediction of tumor recurrence. Compared with the currently widely used optical probes based on intensity signals, the lifetime signal does not depend on the influence of conditions such as the concentration of luminophore, tissue penetration depth and measurement method. Therefore, biological detection methods based on lifetime-based re-sponsive probes have attracted great attention from the scientific community. Here, we briefly review the key advances in lifetime-based responsive probes in recent years (2017)(2018)(2019)(2020)(2021)(2022). The review focuses on the design strategies of lifetime-based responsive probes and the research progress of their applications in the field of bioanalysis, and discusses the challenges they face. We hope it will further promote the development of lifetime-based responsive probes in the field of bioanalysis.

Compact and effective photon-resolved image scanning microscope

“…In fluorescence lifetime ISM (FLISM), 6 , 9 the photon arrival time with respect to the fluorophore excitation event enables mapping the fluorescence lifetime distribution in the sample at superresolution. Such information helps us understand the properties of the biomolecular environment, 18 decipher biomolecule structural changes, 19 and implement multispecies imaging 20 . In quantum ISM (Q-ISM), 5 the photon coincidences allow the construction of an image with a spatial resolution beyond the limits of conventional ISM.…”

Monitoring cancer treatment in live melanoma cells using FLIM

“…However, the use of fluorescent probes for quantitative measurements in conventional fluorescence confocal microscopy presents several limitations such as dependence on fluorophore concentration, excitation source intensity and duration of light exposure, among others [19][20][21]. In contrast, interest in fluorescence lifetime imaging microscopy (FLIM) has increased in the last years because it is both an imaging technique and an absolute measurement method that allows obtaining reproducible concentrations of diverse biological analytes [20,22,23]. Thus, some examples of fluorescence lifetime probes for intracellular pH measurements have been reported in the last decade [24][25][26][27][28][29].…”

Polarity-Sensitive Probe for Two-Photon Fluorescence Lifetime Imaging of Lipid Droplets In Vitro and In Vivo

“…As fluorescence lifetime is an intrinsic property of fluorescence probes, FLIM is not influenced by concentration variations, which can be affected by cell volume changes or photobleaching . Additionally, the sensitivity of fluorescence lifetime to microenvironments enables the visualization and quantification of biophysical parameters such as polarity and viscosity within cells. − Thus, two-photon FLIM (TP-FLIM) is a valuable tool for measuring changes or differences of microenvironments in subcellular regions or in vivo . − To our knowledge, although a few studies have reported the correlation between the polarity change of LDs and the fluorescence lifetime by using one-photon FLIM, , no research has yet reported the polarity differences of LDs in vitro and in vivo by using TP-FLIM. In this study, TBPCPP, a polarity-sensitive two-photon fluorescence lifetime probe for LDs with a donor (D)-acceptor (A)-π-acceptor (A) structure was designed and synthesized by using a facile method (Scheme ).…”