The ability to create complex three-dimensional microstructures has reached an unprecedented level of sophistication in the last 15 years. For the most part, this is the result of a steady development of the additive manufacturing technique named two-photon polymerization (TPP). In a short amount of time, TPP has gone from being a microfabrication novelty employed largely by laser specialists to a useful tool in the hands of scientists and engineers working in a wide range of research fields including microfluidics. When used in combination with traditional microfabrication processes, TPP can be employed to add unique three-dimensional components to planar platforms, thus enabling the realization of lab-on-a-chip solutions otherwise impossible to create. To take full advantage of TPP, an in-depth understanding is required of the materials photochemistry and the fabricated microstructures’ mechanical and chemical properties. Thus, we review methods developed so far to investigate the underling mechanism involved during TPP and analytical methods employed to characterize TPP microstructures. Furthermore, we will discuss potential opportunities for using optofluidics and lab-on-a-chip systems for TPP metrology.
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