NDLI: Yeasts identification in microfluidic devices using peptide nucleic acid fluorescence in situ hybridization (PNA-FISH)

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Yeasts identification in microfluidic devices using peptide nucleic acid fluorescence in situ hybridization (PNA-FISH)

Content Provider	Springer Nature Link
Author	Ferreira, André M. Cruz Moreira, Daniela Cerqueira, Laura Miranda, João M. Azevedo, Nu F.
Copyright Year	2017
Abstract	Peptide nucleic acid fluorescence in situ hybridization (PNA-FISH) is a highly specific molecular method widely used for microbial identification. Nonetheless, and due to the detection limit of this technique, a time-consuming pre-enrichment step is typically required before identification. In here we have developed a lab-on-a-chip device to concentrate cell suspensions and speed up the identification process in yeasts. The PNA-FISH protocol was optimized to target Saccharomyces cerevisiae, a common yeast that is very relevant for several types of food industries. Then, several coin-sized microfluidic devices with different geometries were developed. Using Computational fluid dynamics (CFD), we modeled the hydrodynamics inside the microchannels and selected the most promising options. SU-8 structures were fabricated based on the selected designs and used to produce polydimethylsiloxane-based microchips by soft lithography. As a result, an integrated approach combining microfluidics and PNA-FISH for the rapid identification of S. cerevisiae was achieved. To improve fluid flow inside microchannels and the PNA-FISH labeling, oxygen plasma treatment was applied to the microfluidic devices and a new methodology to introduce the cell suspension and solutions into the microchannels was devised. A strong PNA-FISH signal was observed in cells trapped inside the microchannels, proving that the proposed methodology works as intended. The microfluidic designs and PNA-FISH procedure described in here should be easily adaptable for detection of other microorganisms of similar size.
Starting Page	1
Ending Page	13
Page Count	13
File Format	PDF
ISSN	13872176
Journal	Biomedical Microdevices
Volume Number	19
Issue Number	1
e-ISSN	15728781
Language	English
Publisher	Springer US
Publisher Date	2017-01-31
Publisher Place	New York
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	PNA-Fish Microfluidics Modelling Fluid mechanics Oxygen plasma treatment Biomedical Engineering Biological and Medical Physics, Biophysics Nanotechnology Engineering Fluid Dynamics
Content Type	Text
Resource Type	Article
Subject	Nanoscience and Nanotechnology Molecular Biology Biomedical Engineering

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