No abstract

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.

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.

Recent studies have opened the door to a new generation of photoinitiators for 3D laser nanoprinting. Therein, the simultaneous absorption of two photons, commonly referred to as two‐photon absorption, is replaced by the sequential absorption of two photons in two consecutive one‐photon absorption processes. This process has been termed two‐step absorption. Importantly, two‐step absorption can be accomplished by inexpensive compact low‐power continuous‐wave blue laser diodes instead of femtosecond laser systems in the red spectral region. Red‐shifting the second absorption step with respect to the first one results in an and‐type optical nonlinearity based on two‐color two‐step absorption. Herein, alternatives are systematically explored to the few already reported one‐ and two‐color two‐step‐absorption photoinitiators, including the search for photoinitiators that can be excited by one‐color two‐step absorption and be de‐excited by a disparate laser color.