This application describes some of the salient features of the ensemble of proprietary SPT™ print cartridges we have developed for a variety of purposes. SPT™ print cartridges are compatible with biomaterials and non-biological materials, allowing them to place, position and pattern a broad range of materials on surfaces.

The Nano eNabler™ system is used to directly deliver attoliter to picoliter quantities of an etchant to a gold surface. The result is the production of ultraminiaturized holes in the gold surface, revealing the silicon substrate beneath. This process can be used to create microwells, ultramicrostructures, localized catalytic or inert domains, and ultramicroscale structural elements for building ultraminiaturized devices.

In this application note the Nano eNabler™ system is used to print proteins onto various surface types including hydrogel, nitrocellulose, and chemically treated surfaces such as silanes and self-assembling monolayers (SAMs). This demonstrates the utility of these surfaces as printing substrates for a variety of potential applications, some of which may require a flat silane or SAM surface while others may benefit from the increased loading capacity and activity retention of a 3D matrix. Observation of the printed surfaces provides information on the efficiency of protein transfer and consistency in ultramicron spot formation.

The potential of printed arrays containing multiple proteins in complex or even interlaced
patterns is limitless for the arraying and subsequent monitoring individual, indexed cells. In this
application note, the flexible, multiplexed printing of utilizing the NanoeNabler™ is demonstrated.

Aldehyde-modified 3-aminopropyltriethoxysilane (APTES) can be employed to covalently
bind amines; or, rather, the exposed amino groups of a polypeptide printed with the Nano eNabler™
to borosilicate glass.

Although using the laser for printing arrays controls the “contact force” of the cantilever on the
substrate and this is useful for highly quantitative applications; however, applications, such as printing
arrays for cell binding, do not require a consistent contact force. Because consistent contact force
requires the laser and subsequent associated processing time, printing in “no laser mode” can help
save time in printing large arrays for qualitative applications. We recommend you use a wider cantilever
(30μm or 60μm) for the first attempt.

The standard SPT holder that comes with the Nano eNabler system™ holds the SPT at a 12° angle with respect to the surface. Smaller spots with better morphology and closer spacing can be achieved by using optional, alternate angled SPT holders that hold the cantilever at 24°, 36°, or 48°.  Liquids may also be dispensed into concavities with steeper aspect ratios using these SPT holders.  The one caveat with these holders is that the laser can not be used to find the surface or to print.  We recommend that you use a wider cantilever (30μm or 60μm) for the first attempt.

Epoxysilane SAM (self-assembling monolayers) surfaces on glass or silicon are effective as a substrate for creating protein patterns. Epoxysilanes (glycidoxypropyltrimethoxysilane) can be employed to covalently bind primary amines; and, exposed amino groups of polypeptides printed with the Nano eNabler™ to borosilicate glass.

William Montes and Thomas C. Marsh,  University of St. Thomas, Dept. of Chemistry

The Nano eNabler (NeN) is a versatile micro/nano-scale printing tool for creating arrays of materials with high precision and accuracy.  An open channel microfluidic device is used to deliver very small volumes of solution to a surface.  The majority of current applications for the NeN are focused on creating patterns of water-soluble polymers, biomolecules, viral particles and living cells on various surfaces.  In order to use the NeN for creating arrays of non-polar molecules, a suitable solvent with low vapor pressure is required.  This work describes the development of a sample preparation method and instrument parameters that enable printing arrays of polystyrene (PS) and polymethylmethacrylate (PMMA) onto substrates such as SiO₂, Au, Mica and Indium Tin Oxide.

Indexing living, eukaryotic cells to discrete, addressable loci on surfaces is critical to furthering studies of development, differentiation, response to stimuli, cell-cell communication, regeneration, and in forms of bioelectronics, suturing and tissue replacement. This can be accomplished by employing the Nano eNabler™ to generate arrays of extra-cellular matrix proteins (ECM’s) on APTES-aldehyde, epoxysilane, or polystyrene surfaces. BioForce has successfully indexed NIH3T3, PC12, and HeLa H1 cells.