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• Kim, B
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• Lee, C
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• Park, J
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Sensors 2012, 12(8), 10136-10147; doi:10.3390/s120810136

Article

Synthesis of Bioactive Microcapsules Using a Microfluidic Device

Byeong Il Kim ^1,2,† , Soon Woo Jeong ^1,2,† , Kyoung G. Lee ¹ , Tae Jung Park ³ , Jung Youn Park ⁴ , Jae Jun Song ⁵ , Seok Jae Lee ^1,*  and Chang-Soo Lee ^2,*

¹ Center for Nanobio Integration & Convergence Engineering (NICE), National Nanofab Center, 291 Daehak-ro, Yuseong-gu, Daejeon 305-806, Korea ² Department of Chemical Engineering, Chungnam National University, 220 Gung-Dong, Yuseong-gu, Daejeon 305-764, Korea ³ Department of Chemistry, Chung-Ang University, 84 Heukseok-ro, Dongjak-gu, Seoul 156-756, Korea ⁴ Biotechnology Research Division, National Fisheries Research & Development Institute (NFRDI), 408-1 Sirang-ri, Gijang, Busan 619-705, Korea ⁵ Microbe-based Fusion Technology Research Center, KRIBB, 1404 Sinjeong-dong, Jeongeup, Jeonbuk 580-185, Korea ^† These authors contributed equally to this work.

* Authors to whom correspondence should be addressed.

Received: 8 June 2012; in revised form: 4 July 2012 / Accepted: 18 July 2012 / Published: 26 July 2012

(This article belongs to the Special Issue Live Cell-Based Sensors)

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Abstract: Bioactive microcapsules containing Bacillus thuringiensis (BT) spores were generated by a combination of a hydro gel, microfluidic device and chemical polymerization method. As a proof-of-principle, we used BT spores displaying enhanced green fluorescent protein (EGFP) on the spore surface to spatially direct the EGFP-presenting spores within microcapsules. BT spore-encapsulated microdroplets of uniform size and shape are prepared through a flow-focusing method in a microfluidic device and converted into microcapsules through hydrogel polymerization. The size of microdroplets can be controlled by changing both the dispersion and continuous flow rate. Poly(N-isoproplyacrylamide) (PNIPAM), known as a hydrogel material, was employed as a biocompatible material for the encapsulation of BT spores and long-term storage and outstanding stability. Due to these unique properties of PNIPAM, the nutrients from Luria-Bertani complex medium diffused into the microcapsules and the microencapsulated spores germinated into vegetative cells under adequate environmental conditions. These results suggest that there is no limitation of transferring low-molecular-weight-substrates through the PNIPAM structures, and the viability of microencapsulated spores was confirmed by the culture of vegetative cells after the germinations. This microfluidic-based microencapsulation methodology provides a unique way of synthesizing bioactive microcapsules in a one-step process. This microfluidic-based strategy would be potentially suitable to produce microcapsules of various microbial spores for on-site biosensor analysis.

Keywords: microcapsulation; NIPAM; hydrogel; microfluidic device; spore

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