Two- and three-photon processes during photopolymerization in 3D laser printing.

The performance of a photoinitiator is key to control efficiency and resolution in 3D laser nanoprinting. Upon light absorption, a cascade of competing photophysical processes leads to photochemical reactions toward radical formation that initiates free radical polymerization (FRP). Here, we investigate 7-diethylamino-3-thenoylcoumarin (DETC), belonging to an efficient and frequently used class of photoinitiators in 3D laser printing, and explain the molecular bases of FRP initiation upon DETC photoactivation. Depending on the presence of a co-initiator, DETC causes radical generation either upon two-photon- or three-photon excitation, but the mechanism for these processes is not well understood so far. Here, we show that the unique three-photon based radical formation by DETC, in the absence of a co-initiator, results from its excitation to highly excited triplet states. They allow a hydrogen-atom transfer reaction from the pentaerythritol triacrylate (PETA) monomer to DETC, enabling the formation of the reactive PETA alkyl radical, which initiates FRP. The formation of active DETC radicals is demonstrated to be less spontaneous. In contrast, photoinitiation in the presence of an onium salt co-initiator proceeds via intermolecular electron transfer after the photosensitization of the photoinitiator to the lowest triplet excited state. Our quantum mechanical calculations demonstrate photophysical processes upon the multiphoton activation of DETC and explain different reactions for the radical formation upon DETC photoactivation. This investigation for the first time describes possible pathways of FRP initiation in 3D laser nanoprinting and permits further rational design of efficient photoinitiators to increase the speed and sensitivity of 3D laser nanoprinting.

Two-Photon 3D Laser Printing Inside Synthetic Cells.

Toward the ambitious goal of manufacturing synthetic cells from the bottom up, various cellular components have already been reconstituted inside lipid vesicles. However, the deterministic positioning of these components inside the compartment has remained elusive. Here, by using two-photon 3D laser printing, 2D and 3D hydrogel architectures are manufactured with high precision and nearly arbitrary shape inside preformed giant unilamellar lipid vesicles (GUVs). The required water-soluble photoresist is brought into the GUVs by diffusion in a single mixing step. Crucially, femtosecond two-photon printing inside the compartment does not destroy the GUVs. Beyond this proof-of-principle demonstration, early functional architectures are realized. In particular, a transmembrane structure acting as a pore is 3D printed, thereby allowing for the transport of biological cargo, including DNA, into the synthetic compartment. These experiments show that two-photon 3D laser microprinting can be an important addition to the existing toolbox of synthetic biology.

Two in One: Light as a Tool for 3D Printing and Erasing at the Microscale.

The ability to selectively remove sections from 3D-printed structures with high resolution remains a current challenge in 3D laser lithography. A novel photoresist is introduced to enable the additive fabrication of 3D microstructures at one wavelength and subsequent spatially controlled cleavage of the printed resist at another wavelength. The photoresist is composed of a difunctional acrylate cross-linker containing a photolabile o-nitrobenzyl ether moiety. 3D microstructures are written by photoinduced radical polymerization of acrylates using Ivocerin as photoinitiator upon exposure to 900 nm laser light. Subsequent scanning using a laser at 700 nm wavelength allows for the selective removal of the resist by photocleaving the o-nitrobenzyl group. Both steps rely on two-photon absorption. The fabricated and erased features are imaged using scanning electron microscopy (SEM) and laser scanning microscopy (LSM). In addition, a single wire bond is successfully eliminated from an array, proving the possibility of complete or partial removal of structures on demand.