Journal article
A three-dimensional (3D) printing approach to fabricate an isolation chip for high throughput in situ cultivation of environmental microbes
Lab on a Chip, Vol.22(2), pp.387-402
2022
PMID: 34935836
Abstract
The full plethora of environmental bacteria is often poorly represented in vitro as the majority remain difficult, if not impossible, to culture under standard laboratory settings. These bacteria often require native conditions for the formation of cell masses that collectively have higher chances of survival. With that, a 3D-printed version of the isolation chip (iChip) was used to cultivate bacteria from a tropical peat swamp in situ prior to growth and maintenance in vitro. Briefly, plates made from either acrylonitrile butadiene styrene (ABS), polylactic acid (PLA), or epoxy resin were tested in terms of their usability and durability under acidic conditions similar to those of peat matter. The epoxy resin plates were then found to be most optimal for the sampling conditions. Peat soil samples were collected from the base of a Koompassia malaccensis tree and reconstituted in molten 10% (wt/vol) tryptone soy agar (TSA) prior to inoculation. The iChips were subsequently assembled and buried in the site of origin. As a comparison, bacteria from the same soil sample were cultivated directly on TSA and incubated at 28 °C for two weeks. Thereafter, agar plugs from the iChip were transferred to TSA plates to allow microcolonies within each plug to grow. Each pure isolate from both cultivation approaches that grew was then pooled and extracted for total DNA prior to 16S rRNA gene amplification and sequencing via Illumina MiSeq. Taxonomic abundance comparison revealed that the bacterial taxa at the level of order were significantly different between the two approaches, particularly in the orders, Burkholderiales, Xanthomonodales, Enterobacteriales, and Actinomycetales (differences of 12.0, 7.1, 8.0, and 4.2%, respectively). This indicated that the 3D-printed iChips present a possible low-cost tool for the isolation of bacterial genera that may not be able to grow on media directly in vitro.
Details
- Title
- A three-dimensional (3D) printing approach to fabricate an isolation chip for high throughput in situ cultivation of environmental microbes
- Authors
- Calvin Bok Sun Goh (Author) - Monash University MalaysiaClariss Hui Peng Goh (Author) - Sunway UniversityLi Wen Wong (Author) - Monash University MalaysiaWai Teng Cheng (Author) - Monash University MalaysiaCatherine Mary Yule (Author) - University of the Sunshine Coast, Queensland, School of Science, Technology and EngineeringKuan Shion Ong (Author) - Monash University MalaysiaSui Mae Lee (Author) - Monash University MalaysiaPooria Pasbakhsh (Author) - Monash University MalaysiaJoash Ban Lee Tan (Author) - Monash University Malaysia
- Publication details
- Lab on a Chip, Vol.22(2), pp.387-402
- Publisher
- Royal Society of Chemistry
- DOI
- 10.1039/D1LC00723H
- ISSN
- 1473-0189
- PMID
- 34935836
- Organisation Unit
- University of the Sunshine Coast, Queensland; School of Science, Technology and Engineering
- Language
- English
- Record Identifier
- 99596108302621
- Output Type
- Journal article
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- Domestic collaboration
- International collaboration
- Web Of Science research areas
- Biochemical Research Methods
- Chemistry, Analytical
- Chemistry, Multidisciplinary
- Instruments & Instrumentation
- Nanoscience & Nanotechnology
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