A pincer-like benzene-bridged sensor 1 with a pyrene excimer as a signal source and imidazolium as a phosphate anion receptor was synthesized and investigated for ATP sensing. A unique switch of excimer vs monomer pyrene fluorescence of 1 is observed in the presence of ATP due to the charcteristic sandwich pi-pi stacking of pyrene-adenine-pyrene. On the other hand, four other bases of nucleoside triphosphates such as GTP, CTP, UTP, and TTP can interact only from the outside with the already stabilized stacked pyrene-pyrene dimer of 1, resulting in excimer fluorescence quenching. The fluorescent intensity ratio of monomer-to-excimer for 1 upon binding with ATP (I(375)/I(487)) is much larger than that upon binding with ADP and AMP. This difference is large enough to discriminate ATP from ADP and AMP. As one of the biological applications, sensor 1 is successfully applied to the ATP staining experiments. Sensor 1 is also applied to monitor the hydrolysis of ATP and ADP by apyrase. The results indicate that 1 is a useful fluorescent sensor for investigations of ATP-relevant biological processes.
In this paper we introduce a simple droplet-based microfluidic system consisting of two separate devices to encapsulate and culture microalgae, in contrast to cultivation in bulk liquid medium. This microdroplet technology has been used to monitor the growth of individual microalgal cells in a constant environment for extended periods of time. Single cells from three species of green microalgae, (two freshwater species Chlamydomonas reinhardtii and Chlorella vulgaris, and one saline species Dunaliella tertiolecta), were encapsulated and incubated in microdroplet compartments of diameter of B80 mm, and their growth analysed over 10 days. In all cases, the doubling time of microalgae grown in microdroplets was similar to growth in bulk. The growth of C. reinhardtii in microdroplets of varying diameters and with different initial cell numbers per droplet was investigated, as well as the effect of varying medium conditions such as pH and nitrogen concentration. This methodology offers the opportunity to study characteristics over time of individual cells and colonies, as well as to screen large numbers of them.
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