๐ฃ๐ฎ๐๐๐ฒ๐ฟ๐ป๐ถ๐ป๐ด ๐ฝ๐ฟ๐ผ๐๐ฒ๐ถ๐ป๐ ๐๐ถ๐๐ต ๐น๐ฎ๐๐ฒ๐ฟ-๐ด๐ฟ๐ผ๐๐ป ๐บ๐ถ๐ฐ๐ฟ๐ผ-๐ฏ๐๐ฏ๐ฏ๐น๐ฒ๐: ๐ฎ ๐ฝ๐ผ๐๐๐ถ๐ฏ๐น๐ฒ ๐ป๐ฒ๐ ๐ฟ๐ผ๐๐๐ฒ ๐๐ผ๐๐ฎ๐ฟ๐ฑ๐ ๐น๐ผ๐-๐ฐ๐ผ๐๐ ๐ฑ๐ถ๐ฎ๐ด๐ป๐ผ๐๐๐ถ๐ฐ๐ ๐ฎ๐ป๐ฑ ๐ฏ๐ถ๐ผ๐๐ฒ๐ป๐๐ถ๐ป๐ด
Patterning biomolecules (including proteins such as antigens and antibodies) in pre-designed, parallel architectures can be a game-changer in rapid, inexpensive diagnostics. However, typical patterning methodologies, including top-down approaches such as photolithography and e-beam lithography, or bottom-up approaches such as solgel nanofabrication, molecular self-assembly, and DNA-scaffolding - all have limitations in efficacy for various reasons ranging from damaging the biospecimens to being slow and inflexible.
In this context, Microbubble lithography (MBL) is a comparatively recent (about a decade-old) micro-patterning method that uses laser-generated and translated microbubbles to self-assemble and pattern materials of choice in real time. Thus, MBL has been used to pattern diverse mesoscopic organic and inorganic materials, allowing for various applications including the fabrication of plastic electronics, catalytic chips, and even biosensing. However, in spite of its success in patterning mesoscopic entities, MBL has not yet been employed in developing continuous patterns of living organisms. This is primarily due to the high temperatures associated with a bubble nucleated due to heat generated by tightly focusing a laser โ which is at the heart of this technique.
In this paper, we report the successful creation of a strategy for immobilizing diverse biospecimens on transparent substrates using MBL. Thus, we develop a heterostructured platform made up of a scaffold to form microbubbles, a crosslinker to facilitate adherence, and finally the concerned immobilized biospecimen. Our deployment of fast laser translation speeds ensures no temperature-induced damage to the biospecimens since their contact time with the microbubbles is less than a hundred milliseconds. Our patterned biospecimens cover a wide range of biological macromolecules, such as reporter proteins, bacterial samples, and viruses at varying concentrations. Most importantly, we also performed experiments to determine the activity of the biospecimens, especially bacterial species and virus strains, post patterning โ and obtained conclusive evidence of them retaining their vegetative activity.
Overall, this study sets the groundwork for manufacturing multilayer heterostructures of living matter using MBL, and may open a new paradigm in designing bioelectronic chips, and in fast, inexpensive, and even parallelized biosensing and diagnostics of pathogens, leveraging and highlighting the ubiquitous nature of MBL in terms of patterning โeverything mesoscopicโ.
Article details:
Title: Biologically Active Micropatterns of Biomolecules and Living Matter Using Microbubble Lithography
Authors: Anand Dev Ranjan, Sucharita Bhowmick, Arnab Gupta, Amirul Islam Mallick, Ayan Banerjee
#Research Highlight
Posted on: July 12th, 2024