Lysing Bacterial Spores by Sonication through a Flexible Interface in a Microfluidic System

Michael T. Taylor,* Phillip Belgrader, Burford J. Furman, Farzad Pourahmadi, Gregory T. A. Kovacs,§ and M. Allen Northrup
Cepheid, 1190 Borregas Avenue, Sunnyvale, California 94089, Department of Mechanical & Aerospace Engineering, San Jose State University, San Jose, California 95192, and Stanford University, CISX-202, Stanford, California 94305
Anal. Chem., 2001, 73 (3), pp 492–496
DOI: 10.1021/ac000779v
Publication Date (Web): December 19, 2000
Copyright © 2001 American Chemical Society
*

 Corresponding author. E-mail:  taylor@cepheid.com.

,

 Cepheid.

,

 San Jose State University.

,
§

 Stanford University.

Abstract

Cell disruptions using ultrasonic energy transmitted through a flexible interface into a liquid region has limitations because the motion of the vibrating tip is not completely transferred into the liquid. To ensure that some degree of contact will be maintained between the ultrasonic horn tip and the flexible interface, the liquid must be pressurized. The pressure conditions that yield consistent coupling between the ultrasonic horn tip and the liquid region were explored in this study by using an analytical model of the system and test fixture experiments. The nature of the interaction between the horn tip and the flexible interface creates pulses of positive pressure rises, increase in temperature, streaming flow, and almost no cavitation in the liquid. There was sufficient energy to create a cloud of microspheres, or beads, that maintain a consistent pattern of ballistic motion in the liquid. The sonication was found to be repeatable by studying video recordings of bead motion and was shown to be statistically consistent using measurements of temperature rise. Sonication of bacterial spores to obtain measurements of released nucleic acid and SEM images of damaged spores were used to verify the effects of liquid pressure on the horn−interface−liquid coupling.

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History

  • Published In Issue February 01, 2001
  • Received for review July 7, 2000. Accepted October 25, 2000.

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