Protein diffusion from microwells with contrasting hydrogel domains
Understanding and controlling molecular transport in hydrogel materials is important for biomedical tools, including engineered tissues and drug delivery, as well as life sciences tools for single-cell analysis. Here, we scrutinize the ability of microwells—micromolded in hydrogel slabs—to compartme...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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AIP Publishing LLC
2019-06-01
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| Series: | APL Bioengineering |
| Online Access: | http://dx.doi.org/10.1063/1.5078650 |
| _version_ | 1811192954799259648 |
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| author | Elaine J. Su Shaheen Jeeawoody Amy E. Herr |
| author_facet | Elaine J. Su Shaheen Jeeawoody Amy E. Herr |
| author_sort | Elaine J. Su |
| collection | DOAJ |
| description | Understanding and controlling molecular transport in hydrogel materials is important for biomedical tools, including engineered tissues and drug delivery, as well as life sciences tools for single-cell analysis. Here, we scrutinize the ability of microwells—micromolded in hydrogel slabs—to compartmentalize lysate from single cells. We consider both (i) microwells that are “open” to a large fluid (i.e., liquid) reservoir and (ii) microwells that are “closed,” having been capped with either a slab of high-density polyacrylamide gel or an impermeable glass slide. We use numerical modeling to gain insight into the sensitivity of time-dependent protein concentration distributions on hydrogel partition and protein diffusion coefficients and open and closed microwell configurations. We are primarily concerned with diffusion-driven protein loss from the microwell cavity. Even for closed microwells, confocal fluorescence microscopy reports that a fluid (i.e., liquid) film forms between the hydrogel slabs (median thickness of 1.7 μm). Proteins diffuse from the microwells and into the fluid (i.e., liquid) layer, yet concentration distributions are sensitive to the lid layer partition coefficients and the protein diffusion coefficient. The application of a glass lid or a dense hydrogel retains protein in the microwell, increasing the protein solute concentration in the microwell by ∼7-fold for the first 15 s. Using triggered release of Protein G from microparticles, we validate our simulations by characterizing protein diffusion in a microwell capped with a high-density polyacrylamide gel lid (p > 0.05, Kolmogorov-Smirnov test). Here, we establish and validate a numerical model useful for understanding protein transport in and losses from a hydrogel microwell across a range of boundary conditions. |
| first_indexed | 2024-04-12T00:00:23Z |
| format | Article |
| id | doaj.art-3c5709ab427a4443a01fbf88099f3f71 |
| institution | Directory Open Access Journal |
| issn | 2473-2877 |
| language | English |
| last_indexed | 2024-04-12T00:00:23Z |
| publishDate | 2019-06-01 |
| publisher | AIP Publishing LLC |
| record_format | Article |
| series | APL Bioengineering |
| spelling | doaj.art-3c5709ab427a4443a01fbf88099f3f712022-12-22T03:56:15ZengAIP Publishing LLCAPL Bioengineering2473-28772019-06-0132026101026101-1010.1063/1.5078650001902APBProtein diffusion from microwells with contrasting hydrogel domainsElaine J. Su0Shaheen Jeeawoody1Amy E. Herr2 Department of Bioengineering, University of California, Berkeley, Berkeley, California 94720, USA. Department of Bioengineering, University of California, Berkeley, Berkeley, California 94720, USA. Department of Bioengineering, University of California, Berkeley, Berkeley, California 94720, USA.Understanding and controlling molecular transport in hydrogel materials is important for biomedical tools, including engineered tissues and drug delivery, as well as life sciences tools for single-cell analysis. Here, we scrutinize the ability of microwells—micromolded in hydrogel slabs—to compartmentalize lysate from single cells. We consider both (i) microwells that are “open” to a large fluid (i.e., liquid) reservoir and (ii) microwells that are “closed,” having been capped with either a slab of high-density polyacrylamide gel or an impermeable glass slide. We use numerical modeling to gain insight into the sensitivity of time-dependent protein concentration distributions on hydrogel partition and protein diffusion coefficients and open and closed microwell configurations. We are primarily concerned with diffusion-driven protein loss from the microwell cavity. Even for closed microwells, confocal fluorescence microscopy reports that a fluid (i.e., liquid) film forms between the hydrogel slabs (median thickness of 1.7 μm). Proteins diffuse from the microwells and into the fluid (i.e., liquid) layer, yet concentration distributions are sensitive to the lid layer partition coefficients and the protein diffusion coefficient. The application of a glass lid or a dense hydrogel retains protein in the microwell, increasing the protein solute concentration in the microwell by ∼7-fold for the first 15 s. Using triggered release of Protein G from microparticles, we validate our simulations by characterizing protein diffusion in a microwell capped with a high-density polyacrylamide gel lid (p > 0.05, Kolmogorov-Smirnov test). Here, we establish and validate a numerical model useful for understanding protein transport in and losses from a hydrogel microwell across a range of boundary conditions.http://dx.doi.org/10.1063/1.5078650 |
| spellingShingle | Elaine J. Su Shaheen Jeeawoody Amy E. Herr Protein diffusion from microwells with contrasting hydrogel domains APL Bioengineering |
| title | Protein diffusion from microwells with contrasting hydrogel domains |
| title_full | Protein diffusion from microwells with contrasting hydrogel domains |
| title_fullStr | Protein diffusion from microwells with contrasting hydrogel domains |
| title_full_unstemmed | Protein diffusion from microwells with contrasting hydrogel domains |
| title_short | Protein diffusion from microwells with contrasting hydrogel domains |
| title_sort | protein diffusion from microwells with contrasting hydrogel domains |
| url | http://dx.doi.org/10.1063/1.5078650 |
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