Protein Crosslinking and Immobilization in 3D Microfluidics through Multiphoton Absorption

This study used laser multiphoton absorption (MPA) to crosslink and immobilize protein in 3D microfluidics. MPA can be used to crosslink proteins to form smaller crosslinked-protein structures than those formed through single-photon absorption. However, studies on MPA protein crosslinking in 3D microfluidics are lacking. In this study, the MPA was implemented using pulse laser to induce bovine serum albumin (BSA) crosslinking. We also applied (3-glycidyloxypropyl)trimethoxysilane to increase the adhesion of crosslinked BSA. Also, the width of crosslinked-BSA structure was measured, and then curve-fitting and optical equation were used to predict heights of the crosslinked-BSA structures formed under various laser powers. At last, BSA structures were effectively crosslinked in the three-layered glass or polydimethylsiloxane microchannels. In contrast to studies on single-photon absorption in 3D microfluidics or MPA protein crosslinking in 2D microfluidics, we obtained relatively small crosslinked-BSA structures through MPA and immobilize BSA in 3D microfluidics. In the future, this method can be applied in monolithic 3D microfluidics for protein crosslinking to avoid misalignment and leakage in layer-by-layer 3D microchannels and form more compact 3D microfluidics. These features are conducive to enhance the diversity and convenience of microfluidics in biomedical testing.