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Real-time bacterial microcolony counting using on-chip microscopy

Jung, Jae Hee and Lee, Jung Eun (2016) Real-time bacterial microcolony counting using on-chip microscopy. Scientific Reports, 6 . Art. No. 21473. ISSN 2045-2322. PMCID PMC4763285.

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Observing microbial colonies is the standard method for determining the microbe titer and investigating the behaviors of microbes. Here, we report an automated, real-time bacterial microcolony-counting system implemented on a wide field-of-view (FOV), on-chip microscopy platform, termed ePetri. Using sub-pixel sweeping microscopy (SPSM) with a super-resolution algorithm, this system offers the ability to dynamically track individual bacterial microcolonies over a wide FOV of 5.7 mm × 4.3 mm without requiring a moving stage or lens. As a demonstration, we obtained high-resolution time-series images of S. epidermidis at 20-min intervals. We implemented an image-processing algorithm to analyze the spatiotemporal distribution of microcolonies, the development of which could be observed from a single bacterial cell. Test bacterial colonies with a minimum diameter of 20 μm could be enumerated within 6 h. We showed that our approach not only provides results that are comparable to conventional colony-counting assays but also can be used to monitor the dynamics of colony formation and growth. This microcolony-counting system using on-chip microscopy represents a new platform that substantially reduces the detection time for bacterial colony counting. It uses chip-scale image acquisition and is a simple and compact solution for the automation of colony-counting assays and microbe behavior analysis with applications in antibacterial drug discovery.

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Additional Information:© 2016 Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit Received: 27 September 2015; Accepted: 25 January 2016; Published online: 23 February 2016. We would like to acknowledge Prof. Changhuei Yang (California Institute of Technology) for helpful advice and support on ePetri platform. Also, we acknowledge funding from the KIST Institutional Program. Author Contributions: J.H.J. conceived of the initial idea and designed and implemented the experiment. J.H.J. and J.E.L. developed and refined the concept, and wrote the paper. The authors declare no competing financial interests.
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Korea Institute of Science and Technology (KIST)UNSPECIFIED
PubMed Central ID:PMC4763285
Record Number:CaltechAUTHORS:20160301-091341704
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Official Citation:Jung, J. H. and Lee, J. E. Real-time bacterial microcolony counting using on-chip microscopy. Sci. Rep. 6, 21473; doi: 10.1038/srep21473 (2016).
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:64895
Deposited By: Tony Diaz
Deposited On:01 Mar 2016 23:15
Last Modified:03 Oct 2019 09:42

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