In the past, cellular imaging, and biological analyte assays utilize traditional fluorescence-based techniques. However, since most conventional fluorescence-based probes display only modest fluorescent changes, low sensitivity, low specificity, and so on, those methods have been limited by those drawbacks.1 Recently developed FRET-based probes can overcome those limitations largely to detect and monitor molecular targets in live cells in vitro and in vivo. Moreover, those activatable FRET-based probes can even ratiometriclly measure the concentration of analyte intracellularly or in the tissue sample. Therefore, scientists or clinicians can investigate the specific environmental changes in vitro or diagnostic diseases in time.
Suitable FRET-based probes have been developed to detect H2O2 in mitochondria specifically, DNA methylation in patient tissue samples, and even in vivo imaging with high specificity and more accurate results in bioanalysis, compared to conventional methods.2-3
However, those activatable FRET-based probes are still needed to be improved to make it more valuable in scientific and clinical area, like low toxicity, better cell membrane permeability or even higher sensitivity.4-5
1. Lee, S.; Park, K.; Kim, K.; Choi, K.; Kwon, I. C., Activatable imaging probes with amplified fluorescent signals. Chem Commun 2008, (36), 4250-4260.
2. Ma, Y. F.; Bai, Y. A.; Mao, H. L.; Hong, Q. Y.; Yang, D. W.; Zhang, H. L.; Liu, F. M.; Wu, Z. H.; Jin, Q. H.; Zhou, H. B.; Cao, J.; Zhao, J. L.; Zhong, X. H.; Mao, H. J., A panel of promoter methylation markers for invasive and noninvasive early detection of NSCLC using a quantum dots-based FRET approach. Biosens Bioelectron 2016, 85, 641-648.
3. Du, F. K.; Min, Y. H.; Zeng, F.; Yu, C. M.; Wu, S. Z., A Targeted and FRET- Based Ratiometric Fluorescent Nanoprobe for Imaging Mitochondrial Hydrogen Peroxide in Living Cells. Small 2014, 10 (5), 964-972.
4. Bajar, B. T.; Wang, E. S.; Zhang, S.; Lin, M. Z.; Chu, J., A Guide to Fluorescent Protein FRET Pairs. Sensors-Basel 2016, 16 (9).
5. Rowland, C. E.; Brown, C. W.; Medintz, I. L.; Delehanty, J. B., Intracellular FRET-based probes: a review. Methods Appl Fluores 2015, 3 (4).