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An inexpensive, high-throughput μPAD assay of microbial growth rate and motility on solid surfaces using Saccharomyces cerevisiae and Escherichia coli as model organisms
- Source :
- PLoS ONE, Vol 15, Iss 10, p e0225020 (2020), PLoS ONE
- Publication Year :
- 2020
- Publisher :
- Public Library of Science (PLoS), 2020.
-
Abstract
- Many microbial phenotypes are differentially or exclusively expressed on agar surfaces, including biofilms, motility, and sociality. However, agar-based assays are limited by their low throughput, which increases costs, lab waste, space requirements, and the time required to conduct experiments. Here, we demonstrate the use of wax-printed microfluidic paper-based analytical devices (μPADs) to measure linear growth rate of microbes on an agar growth media as a means of circumventing the aforementioned limitations. The main production materials of the proposed μPAD design are a wax printer, filter paper, and empty pipet boxes. A single wax-printed μPAD allowing 8 independent, agar-grown colonies costs $0.07, compared to $0.20 and $9.37 for the same number of replicates on traditional microtiter/spectrophotometry and Petri dish assays, respectively. We optimized the μPAD design for channel width (3 mm), agar volume (780 μL/channel), and microbe inoculation method (razor-blade). Comparative analyses of the traditional and proposed μPAD methods for measuring growth rate of nonmotile (Saccharomyces cerevisiae) and motile (flagellated Escherichia coli) microorganisms suggested the μPAD assays conferred a comparable degree of accuracy and reliability to growth rate measurements as their traditional counterparts. We substantiated this claim with strong, positive correlations between the traditional and μPAD assay, a significant nonzero slope in the model relating the two assays, a nonsignificant difference between the relative standard errors of the two techniques, and an analysis of inter-device reliability. Therefore, μPAD designs merit consideration for the development of enhanced-throughput, low-cost microbial growth and motility assays.
- Subjects :
- Sucrose
Microfluidics
Yeast and Fungal Models
02 engineering and technology
Bacterial growth
Pathology and Laboratory Medicine
Disaccharides
medicine.disease_cause
01 natural sciences
law.invention
Filter Paper
law
Lab-On-A-Chip Devices
Medicine and Health Sciences
Agar
Materials
Fluids
Multidisciplinary
Microbial Viability
Organic Compounds
Chemistry
Physics
Pipette
Eukaryota
Software Engineering
Equipment Design
Microfluidic Analytical Techniques
021001 nanoscience & nanotechnology
Laboratory Equipment
Experimental Organism Systems
Physical Sciences
Saccharomyces Cerevisiae
Engineering and Technology
Medicine
Fluidics
Pathogens
Organic Materials
0210 nano-technology
Research Article
Pathogen Motility
Paper
Computer and Information Sciences
States of Matter
food.ingredient
Virulence Factors
Science
Materials Science
Carbohydrates
Equipment
Research and Analysis Methods
Models, Biological
Computer Software
Saccharomyces
Model Organisms
food
Escherichia coli
medicine
Chromatography
Filter paper
Petri dish
Organic Chemistry
010401 analytical chemistry
Organisms
Fungi
Chemical Compounds
Biofilm
Biology and Life Sciences
Reproducibility of Results
Liquids
Yeast
0104 chemical sciences
Waxes
Animal Studies
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 15
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....ec91472409ec8f1c00af08842c8728d5