Laser microstructuration of three-dimensional enzyme reactors in microfluidic channels |
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Authors: | Monica Iosin Teodora Scheul Cl??ment Nizak Olivier Stephan Simion Astilean Patrice Baldeck |
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Affiliation: | (1) Nanobiophotonics Laboratory, Institute for Interdisciplinary Experimental Research, Babes-Bolyai University, Treboniu Laurian 42, 400271 Cluj-Napoca, Romania;(2) Laboratoire de Spectrom?trie Physique, Universit? Joseph Fourier, CNRS (UMR 5588), BP 87, 38402 Saint Martin d’H?res Cedex, France |
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Abstract: | In this paper, we report on the fabrication of three-dimensional (3D) enzymatic microreactors within polydimethylsiloxane
microfluidic channels through a photocrosslinking mechanism mediated by the two-photon absorption process at the focal point
of pulse lasers, i.e., a sub-nanosecond Nd:YAG microlaser or a femtosecond Ti:Sapphire laser. This approach allows the building
of localized 3D trypsin structures with submicron resolution. The fabrication of two different trypsin structures was successfully
demonstrated using Eosin Y and Flavin Adenine Dinucleotide as biological photosensitizers: (i) arrays of 3D cylindrical rows
and (ii) 3D woodpile structure. The enzymatic activity of the fabricated structures was evaluated by fluorescence spectroscopy
using BODIPY FL casein as fluorogenic substrate. The real time investigation of the peptide cleavage into the microfluidic
channel demonstrated that the fabricated trypsin microstructures maintain their catalytic activity. This approach opens up
the way to complex multistep enzymatic reactions in well-localized regions of microfluidic devices, with great importance
in health screening and biomedical diagnostics. |
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