Polyacrylamide hydrogels as substrates for studying bacteria

Tuson, Hannah H.; Renner, Lars D.; Weibel, Douglas B.

doi:10.1039/c1cc14705f

Cited by 34 publications

(32 citation statements)

References 35 publications

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“…Longer term experiments, such as used to study aging, would require one to adapt designs including washing channels (see 54,55 for PDMS and [32] for agarose realizations). In addition, polyacrylamide allows chemical modifications for the control of the micro-environment, enabling advanced capabilities such as the controlled release of bacteria [56] or patterned bio-functionalization [57]. …”

Section: Discussionmentioning

confidence: 99%

Microfabricated Polyacrylamide Devices for the Controlled Culture of Growing Cells and Developing Organisms

et al. 2013

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The ability to spatially confine living cells or small organisms while dynamically controlling their aqueous environment is important for a host of microscopy applications. Here, we show how polyacrylamide layers can be patterned to construct simple microfluidic devices for this purpose. We find that polyacrylamide gels can be molded like PDMS into micron-scale structures that can enclose organisms, while being permeable to liquids, and transparent to allow for microscopic observation. We present a range of chemostat-like devices to observe bacterial and yeast growth, and C. elegans nematode development. The devices can integrate PDMS layers and allow for temporal control of nutrient conditions and the presence of drugs on a minute timescale. We show how spatial confinement of motile C. elegans enables for time-lapse microscopy in a parallel fashion.

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Section: Discussionmentioning

confidence: 99%

Microfabricated Polyacrylamide Devices for the Controlled Culture of Growing Cells and Developing Organisms

et al. 2013

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“…As the physical properties of the environment are recognized to play a role in bacterial behavior (24,25,27,28,32), we investigated two factors that may produce differences in cell numbers and growth rates on each substrate. We measured E-the parameter most commonly used to characterize the stiffness of a material-and the diffusion rate of fluorescein-as a surrogate of nutrients-for agar, gellan, xanthan, and BC substrates.…”

Section: Discussionmentioning

confidence: 99%

Bacterial Cellulose as a Substrate for Microbial Cell Culture

Yin

Santos

Auer

et al. 2014

Appl Environ Microbiol

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Bacterial cellulose (BC) has a range of structural and physicochemical properties that make it a particularly useful material for the culture of bacteria. We studied the growth of 14 genera of bacteria on BC substrates produced by Acetobacter xylinum and compared the results to growth on the commercially available biopolymers agar, gellan, and xanthan. We demonstrate that BC produces rates of bacterial cell growth that typically exceed those on the commercial biopolymers and yields cultures with higher titers of cells at stationary phase. The morphology of the cells did not change during growth on BC. The rates of nutrient diffusion in BC being higher than those in other biopolymers is likely a primary factor that leads to higher growth rates. Collectively, our results suggest that the use of BC may open new avenues in microbiology by facilitating bacterial cell culture and isolation.T he introduction of agar as a substrate for bacterial culture in 1882 was an innovation that continues to have a major impact on the microbiology field over a century later (1). A major component of the cell wall of red algae (e.g., the genera Gelidium and Gracilaria), agar consists of monomers of 3,6-anhydro-L-galactose, D-galactose, and L-galactose that are connected by ␤-(1,4) and ␣-(1,3) bonds. The preparation of agar for microbial cell culture involves decreasing the temperature of a solution of agar below its melting temperature (T m ) (typically ϳ50°C), which produces a disordered gel network in which a large volume fraction of water is bound to the hydrophilic polymer chains of agar. Agar has a range of physicochemical characteristics that have made it a widely used material for the culture of microbes, namely, (i) thermal stability, (ii) optical transparency, (iii) biocompatibility, (iv) resistance to degradation by many microorganisms, and (v) availability (2).Agar is by no means an ideal material for all microbiological studies, and its application has been limited by several salient characteristics. First, a number of bacterial species grow poorly or slowly on agar, including Edwardsiella tarda, Klebsiella oxytoca, Klebsiella pneumoniae, Shigella boydii, Shigella flexneri, Staphylococcus saprophyticus, Yersinia enterocolitica, Haemophilus parainfluenzae, Sinorhizobium meliloti, and Bradyrhizobium japonicum (3, 4). Second, agar is a heterogeneous polymer due to variability in the chain length and composition of the polysaccharide. This characteristic can be problematic, as the physicochemical properties of agar vary between manufacturers and batches, which complicates experimental outcomes and reproducibility in phenotypic studies (5, 6). Third, agar is not inert to all bacteria-several genera of bacteria, including Streptomyces, Vibrio, Pseudoalteromonas, Thalassomonas, Alteromonas, Agarivorans, and Microbulbifer, produce and secrete extracellular galactosidases that degrade the hydrogel, which limits its application in bacterial isolation and culture (7). Finally, the costs of harvesting algae and the subsequent proces...

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“…For the creation of bioreactors for the method of additive 3-D technologies the perspective material is gel, in which bioobjects are immobilized. For the cultivation of micro-organisms, agarose gels, and other gels can be used [9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Non-stationary heat transfer in gels applied to biotehnology

et al. 2017

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Unsteady heat transfer in agarose gels of various concentrations was studied in order to make a breakthrough in the technology of 3-D additive bioprinting. Data on the kinetics of the phase transformation was obtained using spectroscopy as a function of temperature during the formation of agarose hydrogel. The dynamics of aging was investigated for gels of different densities. The time dependence of the structural changes was obtained. Particular attention was paid to the changes in the structure of the gel due to the processes of evaporation of the liquid during the gel formation and during long-term storage. Experiments were performed to determine the dynamics of the temperature fields simultaneously with heat flux measurements during the formation of agarose gels from different initial concentrations. A technique based on experimental data for the computations of the thermophysical coefficients of agarose gels was developed.

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Polyacrylamide hydrogels as substrates for studying bacteria

Abstract: Polyacrylamide hydrogels can be used as chemically and physically defined substrates for bacterial cell culture, and enable studies of the influence of surfaces on cell growth and behaviour.

Cited by 34 publications

References 35 publications

Microfabricated Polyacrylamide Devices for the Controlled Culture of Growing Cells and Developing Organisms

Microfabricated Polyacrylamide Devices for the Controlled Culture of Growing Cells and Developing Organisms

Bacterial Cellulose as a Substrate for Microbial Cell Culture

Non-stationary heat transfer in gels applied to biotehnology

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