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Ship-in-a-bottle integration by hybrid femtosecond laser technology for fabrication of true 3D biochips

We propose herein the “ship-in-a-bottle” integration of three-dimensional (3D) polymeric sinusoidal ridges inside photosensitive glass microfluidic channel by a hybrid subtractive - additive femtosecond laser processing method. It consists of Femtosecond Laser Assisted Wet Etching (FLAE) of a photosensitive Foturan glass followed by Two-Photon Polymerization (TPP) of a SU-8 negative epoxy-resin. Both subtractive and additive processes are carried out using the same set-up with the change of laser focusing objective only. A 522 nm wavelength of the second harmonic generation from an amplified femtosecond Yb-fiber laser (FCPA µJewel D-400, IMRA America, 1045 nm; pulse width 360 fs, repetition rate 200 kHz) was employed for irradiation. The new method allows lowering the size limit of 3D objects created inside channels to smaller details down to the dimensions of a cell, and improve the structure stability. Sinusoidal periodic patterns and ridges are of great use as base scaffolds for building up new structures on their top or for modulating cell migration, guidance and orientation while created interspaces can be exploited for microfluidic applications. The glass microchannel offers robustness and appropriate dynamic flow conditions for cellular studies while the integrated patterns are reducing the size of structure to the level of cells responsiveness. Taking advantage of the ability to directly fabricate 3D complex shapes, both glass channels and polymeric integrated patterns enable us to 3D spatially design biochips for specific applications.