nano3D printer

nano3D printer: The nano3D printer was invented by Dr. Felix Löffler. This robot was developed within the BMBF project KATMETHAN and EU projects PEPDIODE (more) and TARGETBINDER (more). Currently, we advance it together with Dr. Dario Mager, Dr. Felix Löffler, and SME PEPperPRINT to an industry-robot with a Technology Readiness Level of 9 (TRL9).

Synthesis of peptide arrays with the nano3D printer: The nano3D printer uses single laser pulses to transfer “small amino-acid-spots” from a donor to an acceptor. [8; see also Fig. 5] Simply by consecutively exchanging the first donor slide with additional donor slides the acceptor is structured with all the 20 different amino acid building blocks – each embedded within the same solid material that is also used for the “peptide-laser-printer” (more) and the “one-cavity-one-peptide-method”. (more)

The nano3D printer does the printing with the help of single laser pulses that are directed to selected spots on a surface via a 2D laser scanning system: These pulses are absorbed by a polyimide foil, and, thereby, “translated” into a heatwave that travels to the surface of the donor foil until it reaches the interphase to the surrounding air, where it melts a few molecule layers. At the same time, the heat-absorbing polyimide expands and forms a bubble that lightly touches the acceptor slide underneath the donor slide. [11] Thereby, and similar to a humid finger that touches a very cold metal piece, a few molecule layers are frozen to the acceptor slide (Figs. 4 & 5).

Fig. 4; nano3D printer, printing mode. Left) A single laser pulse is absorbed by the polyimide layer, leading to a reversible expansion of the polyimide that lightly touches the acceptor glass slide underneath. Right) Schematically shown is the expanding polyimide foil with its to-be-printed-material that was metered onto the polyimide. Simultaneous to expanding the polyimide layer, the heatwave melts a few molecule layers of the to-be-printed-material at the interface to the surrounding air. These are frozen as a nanolayer to the acceptor surface. 

Fig. 5; nano3D printer. A) View of the nano-3D-printer with its different functional units (AOM = acousto-optic modulator). B) A coated donor slide is positioned over an acceptor substrate. Next, single laser pulses from a 2D laser scanning system “punch out” tiny material spots that are transferred to the acceptor slide. Simply by repeating this processing step with additional donor slides leads to the structuring of the acceptor slide with many different nanolayers that comprise polymer-embedded, and, thereby, stabilized chemicals. These solid nanolayers are stacked by the robot in freely chosen combinations and stoichiometry. C) When melted by solvent vapor in a controlled atmosphere, extremely miniaturized chemical reactions are started in the array format. Removal of solvent vapor stops these chemical reactions, and additional chemicals can be added to do multistep one-pot reactions. MALDI MS imaging can be used to analyse reaction products. D) Transferred material spots are nanoscale (measured with vertical scanning interferometry). The transfer of a first, a second, a third, and a fourth layer of matrix material (the material that is used to embed Fmoc-amino-acid building blocks) is shown. Stacked layers from 1×, 2×, 3×, 4× printing are depicted in different colors to highlight added material.

Figure from Mattes et al., Advanced Materials, first online published 29 April 2019; 31: 1806656; DOI: 10.1002/adma.201806656

Technical advantages of the nano3D printer are (i) very small spot sizes due to structuring with solid materials, (ii) stability of reactive chemicals due to shielding then within a solid material, and (iii) the “ready-to-go-status” of polymer-embedded chemical reactants – once melted, they immediately start to diffuse towards their reaction partner. [9] Another big advantage with respect to peptide array synthesis is that the nano3D printer – and the one-cavity-one-peptide-method – can easily synthesize peptide arrays with posttranslational modified amino acids. [8] Thereby, we think, completely new applications for peptide arrays should be feasible (more).

However, recently we learned the nano3D printer’s novel feature of multi-material combinatorial printing of nanolayers lends itself also to applications for peptide arrays applications beyond peptide arrays synthesis: [9] funded by the FET-Proactive proposal NANOSTACKS (more) we explore new opportunities in materials research (more) and in combinatorial extremely miniaturized chemical synthesis in the array format. (more)

[8] Loeffler FF, Foertsch TC, Popov R, Mattes DS, Schlageter M, Sedlmayr M, Ridder B, Dang FX, von Bojničić-Kninski C, Weber LK, Fischer A,Greifenstein J, Bykovskaya V, Buliev I, Bischoff FR, Hahn L, Meier MAR, Bräse S, Powell AK, Balaban TS, Breitling F, Nesterov-Mueller A. High-flexibility combinatorial peptide synthesis with laser-based transfer of monomers in matrix material. Nature Communications, 2016, DOI: 10.1038/NCOMMS11844

[9] Mattes DS, Jung N, Weber L, Bräse S, Breitling F. (2019) Miniaturized and automated synthesis of biomolecules – Overview and perspectives. Advanced Materials, first online published 29 April 2019; 31: 1806656; DOI: 10.1002/adma.201806656

[10] Stadler V, Kirmse R, Beyer M, Breitling F, Ludwig T, and Bischoff FR. (2008) PEGMA/MMA Copolymer Graftings: Generation, Protein Resistance, and a Hydrophobic Domain. Langmuir 24, 8151-8157

[11] Paris G, Klinkusch A, Heidepriem J, Tsouka A, Zhang J, Mende M, Mattes DS, Mager D, Riegler H, Eickelmann S, Loeffler FF. (2020) Laser-induced forward transfer of soft material nanolayers with millisecond pulses shows contact-based material deposition. Applied surface science, 508, Art. Nr.: 144973. DOI: 10.1016/j.apsusc.2019.144973