Direct-write patterning of bacterial cells by dip-pen nanolithography

Area of Science:

• Biotechnology
• Nanotechnology
• Microbiology

Background:

• Dip-pen nanolithography (DPN) excels at creating nano/microarrays of various materials.
• Transporting large materials like bacteria with DPN is challenging due to water meniscus size limitations.

Purpose of the Study:

• To develop a straightforward "stamp-on" DPN method for patterning micrometer-sized bacterial cells.
• To overcome the size limitations of traditional DPN for biological material transport.

Main Methods:

• Utilized a nanostructured poly(2-methyl-2-oxazoline) hydrogel-coated tip with carrier agents.
• Employed a "stamp-on" DPN approach for direct-write patterning of Escherichia coli JM 109 cells.
• Modulated ink solution viscosity to control deposition of single cells or multiple layers.

Main Results:

• Successfully generated patterns of micrometer-sized Escherichia coli JM 109 bacterial cells.
• Demonstrated deposition of single bacterial cell arrays and multi-layer arrays.
• Confirmed bacterial viability post-DPN patterning on Luria-Bertani-agar surfaces via fluorescence microscopy.

Conclusions:

• The novel "stamp-on" DPN method effectively overcomes size limitations for patterning live bacterial cells.
• This technique offers precise control over bacterial cell array generation, from single cells to multi-layer structures.
• The method preserves bacterial viability, opening possibilities for advanced bio-patterning applications.