Freeform multiphoton excited microfabrication for biological applications using a rapid prototyping CAD-based approach

Abstract

Multiphoton excited polymerization has attracted increasing attention as a powerful 3 dimensional nano/microfabrication tool. The nonlinear excitation confines the fabrication region to the focal volume allowing the potential to achieve freeform fabrication with submicron capabilities. We report the adaptation and use of a computer aided design (CAD) approach, based on rapid prototyping software, which exploits this potential for fabricating with protein and polymers in biologically compatible aqueous environments. 3D structures are drawn in the STL format creating a solid model that can be sliced, where the individual sections are then serially fabricated without overwriting previous layers. The method is shown for potential biological applications including microfluidics, cell entrapment, and tissue engineering.

Spatial and Temporal Analysis of Extracellular Matrix Proteins in the Developing Murine Heart: A Blueprint for Regeneration

“…We demonstrated that this spatial information can be used in conjunction with a controlled fabrication technique to create a synthetic scaffold that imitates the organization of the COLIV matrix in the postnatal day 2 mouse ventricle. Because this fabrication technology can be utilized with ECM proteins, 50 this methodology could create ECM protein scaffolds mimicking the in vivo microenvironment of the heart, or other region of interest, during a developmental stage at which the microenvironment elicits cues to its occupying cells to perform a desired function. For example, research groups interested in maintaining, propagating, and encouraging differentiation of cardiac progenitors might consider utilizing the ECM blueprint defined at E16.5.…”

“…50 The same region was imaged in 20 serial sections from a postnatal day 2 ventricle labeled with the anti-COLIV antibody. Antibody labeling and subsequent imaging were performed as described above.…”

“…This scaffold was generated via Computer Aided Design (CAD) control coupled to multiphoton excitation (MPE) fabrication 50 of TMPTA. To accomplish this, the same region was imaged in 20 serial sections (*100 mm total thickness) from a day 2 postnatal ventricle labeled with the COLIV antibody.…”

Two-photon polymerization of hydrogels – versatile solutions to fabricate well-defined 3D structures

“…72,73 Because Rose Bengal displayed a low solubility at acidic pH, where collagens have the highest solubility, another photoinitiator that is soluble at acidic pH was considered. Consequently, they reported a newly synthesized benzophenone dimer that could act as a photoinitiator for both BSA and collagen cross-linking.…”

Simultaneous additive and subtractive three-dimensional nanofabrication using integrated two-photon polymerization and multiphoton ablation

“…In recent years, three-dimensional (3D) micro-/nanofabrication techniques have attracted increasing attention, due to their promise in a wide range of applications including integrated optics, 1,2 plasmonics, 3,4 microbiology 5,6 and microelectromechanical systems. [7][8][9][10][11] Similar to 3D fabrication at macroscopic scales, 3D micro-/nanofabrication generally entails two fundamental approaches: additive and subtractive processes.…”