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Photodegradable Hydrogels for Cell Release and Retrieval

Retrieval of cells from interfaces after microscopic observation for follow-up “omics” measurements is a critical but often overlooked step that limits many cell-based assays. In this work, photodegradable hydrogels are developed to non-destructively recover cells from screening interfaces in order to couple an observed phenotype with genomic information. Polyethylene glycol (PEG)-based hydrogels are used to first immobilize individual bacteria cells from a heterogenous population for culture and/or microscopic observation. PEG linkers are modified with photo-cleavable o-nitrobenzyl chromophores, allowing for selective degradation of hydrogel areas with high spatial precision using a patterned illumination tool and a 365 nm LED light source. Light exposure triggers controlled hydrogel degradation and efficient release of targeted bacteria cells into solution for retrieval, enrichment, and sequencing. Using this methodology, photodegradable hydrogels have been developed as a light-responsive membrane for use on a microwell co-culture platform. Hydrogels are currently developed towards isolation and characterization at the single level in effort to discover cells with rare or unique function. 

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Isolation of a phenotypically rare microbial cell from a photodegradable hydrogel cell culture matrix using micropatterned light. 

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Publications:

 

  • M. M. Gutierrez, J.A. Reed, R. A. McElroy, R.R. Hansen, “Bacteria Encapsulation into Polyethylene Glycol Hydrogels Using Michael-Type Addition Reactions”, MRS Advances (2024). doi: 10.1557/s43580-024-00940-y

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  • M. Masigol, E.L. Radaha, A. Kannan, A.G. Salberg, N. Fattahi, P. Parameswaran, R.R. Hansen, "Polymer Surface Dissection for Correlated Microscopic and Compositional Analysis of Bacterial Aggregates During Membrane Biofouling", ACS Applied Bio Materials, 5(1) 134-145 (2022). doi: 10.1021/acsabm.1c00971    

                                     

  • R.R. Hansen, T.G. Platt, A.J. van der Vlies. “Hydrogel Membrane and Methods for Selective Retrieval of Microbial Targets”, U.S. Pat. No. 11,235,330, 2022.

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  • N. Fattahi, N. Barua, A.J. van der Vlies, R.R. Hansen, "Photodegradable Hydrogel Interfaces for Bacteria Screening, Selection, and Isolation", Journal of Visualized Experiments, 177, e63048 (2021). doi: 10.3791/63048

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  • N. Fattahi, P. Guzman, M. Masigol, A. J. van der Vlies, R. Jensen, R.R. Hansen, and T.G. Platt, “Photodegradable hydrogels for rapid screening, isolation, and genetic characterization of bacteria with rare phenotypes”, Biomacromolecules, 21(8) 3140-3151 (2020). doi: 10.1021/acs.biomac.0c00543

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  • A. J. van der Vlies, N. Barua, P.A. Nieves-Otero, T.G. Platt, and R.R. Hansen, “On Demand Release and Retrieval of Bacteria from Microwell Arrays Using Photodegradable Hydrogel Membranes”, ACS Applied Bio Materials, 2(1), 266-276 (2019). doi: 10.1021/acsabm.8b00592

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