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A three-dimensional (3D) printing approach to fabricate an isolation chip for high throughput in situ cultivation of environmental microbes
Journal article   Peer reviewed

A three-dimensional (3D) printing approach to fabricate an isolation chip for high throughput in situ cultivation of environmental microbes

Calvin Bok Sun Goh, Clariss Hui Peng Goh, Li Wen Wong, Wai Teng Cheng, Catherine Mary Yule, Kuan Shion Ong, Sui Mae Lee, Pooria Pasbakhsh and Joash Ban Lee Tan
Lab on a Chip, Vol.22(2), pp.387-402
2022
DOI: https://doi.org/10.1039/d1lc00723h
PMID: 34935836
url
https://doi.org/10.1039/D1LC00723HView
Published Version

Abstract

peat swamp forest Polylactic acid soil bacteria pure culture deposition blends polycarbonate culturability biodiversity antibiotics
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.

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Collaboration types
Domestic collaboration
International collaboration
Web Of Science research areas
Biochemical Research Methods
Chemistry, Analytical
Chemistry, Multidisciplinary
Instruments & Instrumentation
Nanoscience & Nanotechnology

UN Sustainable Development Goals (SDGs)

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#15 Life on Land

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