The peptide sequences of the four FRET-based substrates ([..] CPC Scientific) are as follows: UV: AlexaFluor405-Leu-Ala-Gln-Ala-HompheArg-Ser-Lys (QSY35)-NH2; Blue: Dabcyl-Gly-Pro-Leu-Gly-Met-Arg-Gly-Lys (5-FAM)-NH2; Green: QSY7-Ala-Pro-Phe-Glu..
Abstract
The recent maturation of continuous-flow microfluidic technologies has coincided with transformative new methods to profile single cells, including their genetic types, protein expression and enzyme activities. Continuous-flow high-throughput single-cell screening and sorting can reveal relationships across cellular phenotypes (e.g., enzyme activity and secretion) and genetic fingerprints. This technology provides unique opportunities, as well as experimental and computational challenges, for integrative approaches that can process large amounts of single-cell data. In this chapter, we discuss recent advances in integrated continuous-flow microfluidic approaches with a focus on measurements and statistical analysis of single-cell enzyme activity and their applications in quantitative biology, synthetic biology, and diagnosis.
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