Issue 4, 2012
From the journal:

Lab on a Chip

Sonolysis of Escherichia coli and Pichia pastoris in microfluidics

Tandiono Tandiono,†*a   Dave Siak-Wei Ow,†b   Leonie Driessen,c   Cara Sze-Hui Chin,b   Evert Klaseboer,a   Andre Boon-Hwa Choo,b   Siew-Wan Ohla  and  Claus-Dieter Ohl*c  

* Corresponding authors

a Institute of High Performance Computing, 1 Fusionopolis Way, #16-16 Connexis, Singapore, Singapore
E-mail: tandiono@ihpc.a-star.edu.sg

b Bioprocessing Technology Institute, 20 Biopolis Way, #06-01 Centros, Singapore, Singapore

c Division of Physics and Applied Physics, School of Physical and Mathematical Sciences, Nanyang Technological University, Singapore, Singapore
E-mail: cdohl@ntu.edu.sg

Abstract

We report on an efficient ultrasound based technique for lysing Escherichia coli and Pichia pastoris with oscillating cavitation bubbles in an integrated microfluidic system. The system consists of a meandering microfluidic channel and four piezoelectric transducers mounted on a glass substrate, with the ultrasound exposure and gas pressure regulated by an automatic control system. Controlled lysis of bacterial and yeast cells expressing green fluorescence protein (GFP) is studied with high-speed photography and fluorescence microscopy, and quantified with real-time polymerase chain reaction (qRT-PCR) and fluorescence intensity. The effectiveness of cell lysis correlates with the duration of ultrasound exposure. Complete lysis can be achieved within one second of ultrasound exposure with a temperature increase of less than 3.3 °C. The rod-shaped E. coli bacteria are disrupted into small fragments in less than 0.4 seconds, while the more robust elliptical P. pastoris yeast cells require around 1.0 second for complete lysis. Fluorescence intensity measurements and qRT-PCR analysis show that functionality of GFP and genomic DNA for downstream analytical assays is maintained.

Graphical abstract: Sonolysis of Escherichia coli and Pichia pastoris in microfluidics

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Article information

DOI
https://doi.org/10.1039/C2LC20861J
Article type
Paper
Submitted
09 Sep 2011
Accepted
28 Nov 2011
First published
20 Dec 2011

Lab Chip, 2012,12, 780-786

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Sonolysis of Escherichia coli and Pichia pastoris in microfluidics

T. Tandiono, D. Siak-Wei Ow, L. Driessen, C. Sze-Hui Chin, E. Klaseboer, A. Boon-Hwa Choo, S. Ohl and C. Ohl, Lab Chip, 2012, 12, 780 DOI: 10.1039/C2LC20861J

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