Automated single cell microbioreactor for monitoring intracellular dynamics and cell growth in free solution.
We report an automated microfluidic-based platform for single cell analysis that allows for cell culture in free solution with the ability to control the cell growth environment. Using this approach, cells are confined by the sole action of gentle fluid flow, thereby enabling non-perturbative analysis of cell growth away from solid boundaries. In addition, the single cell microbioreactor allows for precise and time-dependent control over cell culture media, with the combined ability to observe the dynamics of non-adherent cells over long time scales. As a proof-of-principle demonstration, we used the platform to observe dynamic cell growth, gene expression, and intracellular diffusion of repressor proteins while precisely tuning the cell growth environment. Overall, this microfluidic approach enables the direct observation of cellular dynamics with exquisite control over environmental conditions, which will be useful for quantifying the behaviour of single cells in well-defined media.
Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Eric M
EM
Johnson-Chavarria
Utsav
U
Agrawal
Melikhan
M
Tanyeri
Thomas E
TE
Kuhlman
Charles M
CM
Schroeder
D057205:Automation, Laboratory; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D019149:Bioreactors / Q000382:microbiology*; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004867:Equipment Design; D004926:Escherichia coli / Q000254:growth & development / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D008422:Materials Testing; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008856:Microscopy, Fluorescence; D018715:Microscopy, Video; D055668:Photochemical Processes; D017319:Photosensitizing Agents / Q000737:chemistry; D066330:Printing, Three-Dimensional; D021381:Protein Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism; D059010:Single-Cell Analysis / Q000295:instrumentation*; D013499:Surface Properties; D059008:Time-Lapse Imaging
D001426:Bacterial Proteins; D004129:Dimethylpolysiloxanes; D029968:Escherichia coli Proteins; D008164:Luminescent Proteins; D017319:Photosensitizing Agents; D011994:Recombinant Proteins; D012097:Repressor Proteins; C054562:yellow fluorescent protein, Bacteria; C013830:baysilon