1. Fluorescence Anisotropy as a Temperature-Sensing Molecular Probe Using Fluorescein
- Author
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Puneet Jain, Takuya Aida, and Masahiro Motosuke
- Subjects
fluorescence anisotropy ,fluorescein ,microfluidics ,fluorescence polarization ,temperature imaging ,thermal molecular probe ,Mechanical engineering and machinery ,TJ1-1570 - Abstract
Fluorescence anisotropy, a technique to study the folding state of proteins or affinity of ligands, is used in this present work as a temperature sensor, to measure the microfluidic temperature field, by adding fluorophore in the liquid. Fluorescein was used as a temperature-sensing probe, while glycerol–aq. ammonia solution was used as a working fluid. Fluorescence anisotropy of fluorescein was measured by varying various parameters. Apart from this, a comparison of fluorescence anisotropy and fluorescence intensity is also performed to demonstrate the validity of anisotropy to be applied in a microfluidic field with non-uniform liquid thickness. Viscosity dependence and temperature dependence on the anisotropy are also clarified; the results indicate an appropriate selection of relation between molecule size and viscosity is important to obtain a large temperature coefficient in anisotropy. Furthermore, a practical calibration procedure of the apparatus constant is proposed. In addition, the potential of temperature imaging is confirmed by the measurement of temperature distribution under focused laser heating.
- Published
- 2021
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