A Caltech Library Service

Propidium monoazide pretreatment on a 3D-printed microfluidic device for efficient PCR determination of 'live versus dead' microbial cells

Zhu, Yanzhe and Huang, Xiao and Xie, Xing and Bahnemann, Janina and Lin, Xingyu and Wu, Xunyi and Wang, Siwen and Hoffmann, Michael R. (2018) Propidium monoazide pretreatment on a 3D-printed microfluidic device for efficient PCR determination of 'live versus dead' microbial cells. Environmental Science: Water Research & Technology, 4 (7). pp. 956-963. ISSN 2053-1400. PMCID PMC7705123.

[img] PDF - Published Version
Creative Commons Attribution.

[img] PDF - Supplemental Material
Creative Commons Attribution.


Use this Persistent URL to link to this item:


Waterborne microbial pathogen detection via nucleic acid analysis on portable microfluidic devices is a growing area of research, development, and application. Traditional polymerase chain reaction (PCR)-based nucleic acid analysis detects total extracted DNA, but cannot differentiate live and dead cells. A propidium monoazide (PMA) pretreatment step before PCR can effectively exclude DNA from nonviable cells, as PMA can selectively diffuse through compromised cell membranes and intercalate with DNA to form DNA–PMA complex upon light exposure. The complex strongly inhibits the amplification of the bound DNA in PCR, and thus, only cells with intact cell membranes are detected. Herein, this study reports the development of a microfluidic device to carry out PMA pretreatment ‘on-chip’. Chip design was guided by computer simulations, and prototypes were fabricated using a high-resolution 3D printer. The optimized design utilizes split and recombine mixers for initial PMA-sample mixing and a serpentine flow channel containing herringbone structures for dark and light incubation. On-chip PMA pretreatment to differentiate live and dead bacterial cells in buffer and natural pond water samples was successfully demonstrated.

Item Type:Article
Related URLs:
URLURL TypeDescription Information CentralArticle
Zhu, Yanzhe0000-0002-2260-1830
Huang, Xiao0000-0002-3737-6939
Xie, Xing0000-0002-2253-0964
Bahnemann, Janina0000-0002-7008-1673
Lin, Xingyu0000-0002-0950-0736
Wang, Siwen0000-0002-8553-425X
Hoffmann, Michael R.0000-0001-6495-1946
Additional Information:© 2018 The Royal Society of Chemistry. Open Access Articles. Published on 11 June 2018. This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. The article was received on 31 Jan 2018, accepted on 11 Apr 2018 and first published on 11 Jun 2018. The authors acknowledge the financial support of the Bill and Melinda Gates Foundation (BMGF OPP1111246). There are no conflicts to declare.
Funding AgencyGrant Number
Bill and Melinda Gates FoundationOPP1111246
Issue or Number:7
PubMed Central ID:PMC7705123
Record Number:CaltechAUTHORS:20180611-134345584
Persistent URL:
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:86966
Deposited By: Tony Diaz
Deposited On:11 Jun 2018 20:54
Last Modified:04 Jan 2021 18:37

Repository Staff Only: item control page