Abstract:
The dataset includes microchip design tests to later investigate the effect of oil, Corexit or both on the growth and production of a marine microbe species.
Suggested Citation:
James Chiu, Wei-Chun Chin. 2017. Microbial species growth and exopolymeric substance production microchip screening test. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N78C9TCZ
Publications:
Chiu, M.-H., Vazquez, C. I., Shiu, R.-F., Le, C., Sanchez, N. R., Kagiri, A., … Chin, W.-C. (2019). Impact of exposure of crude oil and dispersant (Corexit) on aggregation of extracellular polymeric substances. Science of The Total Environment, 657, 1535–1542. doi:10.1016/j.scitotenv.2018.12.147
Shiu, R.-F., Vazquez, C. I., Tsai, Y.-Y., Torres, G. V., Chen, C.-S., Santschi, P. H., … Chin, W.-C. (2020). Nano-plastics induce aquatic particulate organic matter (microgels) formation. Science of The Total Environment, 706, 135681. doi:10.1016/j.scitotenv.2019.135681
Shiu, R.-F., Chiu, M.-H., Vazquez, C. I., Tsai, Y.-Y., Le, A., Kagiri, A., … Chin, W.-C. (2020). Protein to carbohydrate (P/C) ratio changes in microbial extracellular polymeric substances induced by oil and Corexit. Marine Chemistry, 223, 103789. doi:10.1016/j.marchem.2020.103789
Purpose:
Testing a microfluidics design for microbial EPS production.
Data Parameters and Units:
A video of the microfluidics of a chip: Shows gradients generate by food dye in our microfluid device. Blue dye is an analog for a chemical, yellow represents a marine organism. When the blue dye (chemical) diffuses from the lower side to upper side into the yellow dye (marine organism), this represents the chemical moving from a high concentration to a low concentration, over a time span of 24 hrs. B1 Material and Method_explanation of the microfluidic chip B2 Figure 1A: screen shot from the video to show what the device looks like. B3 Figure 1B: Co-Draw Design software file showing the device pattern. B4 Figure 1C: picture of the device with green dye added in chemical well after 18 hrs and 24 hrs. C Design 1: first design of microfluid device D Design 2: new design with enlarge sample well in microfluid device E Modified chip design 1: Simplified version 1 to teach outreach groups F Modified chip design 2: Simplified version 2 to teach outreach groups G Model of the microfluidics of a chip: Shows gradients mixing in a cross- sectional view of the flow
Methods:
Microfluid device. Acrylics were purchased from McMaster.com and cut by UNIVERSAL LASER SYSTEM VLS2.30 for particular patterns designed using Co-Draw Design software. The bottom of the semi-finished material was sealed using Polydimethylsiloxane (PDMS) in a petri-dish, and vacuum overnight treatment and then dry by 65 °C for another overnight. Nanoparticle diffusion. Nanoparticles (NP) of different sizes (53nm, 250nm, 500nm) were conjugated with dragon green (480nm/520nm) florescent dye purchased from Bangs Laboratories Inc. These were used in for diffusion measurements in the microfluidics device. Particles were diluted in L1 medium, an enriched algal growth medium, to make in 0.05% concentration. Dextran (Sigma®) was diluted by L1 medium to make 12.5% to control nanoparticle diffusion efficiency. PerkinElmer ELISA reader VICTOR3 was used to read nanoparticle florescence intensity. Gradients within microfluidics. Microfluidic devices were successful used to generate concentration gradients to accelerate the emergence of resistance mutants (Frisch & Rosenberg, 2011; Zhang et al., 2011). In this study, we propose to build a microfluidic device that is for monitor toxicity of gradient concentration of NPs (nanoparticles), oil, or dispersant to diatoms. Figure 1A shows the microfluidic design with dyes to simulate the diffusion within the channels of the device. Figure 1B shows the design followed by the laser cutter. In Figure 1C, diatoms were grown in the channel (represented as yellow pigment). The green pigment represents the NPs, oil or Corexit. In fig. 1 B, diatoms can be collected from round wells 1 to 7, to measure the different NP size concentrations for following experiment. These NP concentrations were measured using 480nm excitation wavelength for the NPs and observed through the fluorescence reader, at the 520 nm emission wavelength as a percentage of the gradient concentration in relative fluorescence. We also add dextran, which is a polymer as a blocker, to control the NPs diffusion. In Figure1C, 12.5% dextran was added to NPs which reduced the NPs diffusion efficiency down to 50%. Therefore, dextran is a good diffusion regulator in this device for future experiment.
Provenance and Historical References:
Frisch RL, & Rosenberg SM (2011). Microbiology. Antibiotic resistance, not shaken or stirred. Science 333: 1713-1714. Shi H, Magaye R, Castranova V, & Zhao J (2013). Titanium dioxide nanoparticles: a review of current toxicological data. Particle and fibre toxicology 10: 15. Zhang Q, Lambert G, Liao D, Kim H, Robin K, Tung CK, et al. (2011). Acceleration of emergence of bacterial antibiotic resistance in connected microenvironments. Science 333: 1764-1767. Zhang SJ, Jiang YL, Chen CS, Creeley D, Schwehr KA, Quigg A, et al. (2013). Ameliorating effects of extracellular polymeric substances excreted by Thalassiosira pseudonana on algal toxicity of CdSe quantum dots. Aquat Toxicol 126: 214-223.
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