Dataset Information

Characterization of Tensioned PDMS Membranes for Imaging Cytometry on Microraft Arrays.

ABSTRACT: Polydimethylsiloxane (PDMS) membranes can act as sensing elements, barriers, and substrates, yet the low rigidity of the elastomeric membranes can limit their practical use in devices. Microraft arrays rely on a freestanding PDMS membrane as a substrate for cell arrays used in imaging cytometry and cellular isolation. However, the underlying PDMS membrane deforms under the weight of the cell media, making automated analytical microscopy (and thus cytometry and cell isolation) challenging. Here we report the development of microfabrication strategies and physically motivated mathematical modeling of membrane deformation of PDMS microarrays. Microraft arrays were fabricated with mechanical tension stored within the PDMS substrate. These membranes deformed 20× less than that of arrays fabricated using prior methods. Modeling of the deformation of pretensioned arrays using linear membrane theory yielded ?15% error in predicting the array deflection and predicted the impact of cure temperatures up to 120 °C. A mathematical approach was developed to fit models of microraft shape to sparse real-world shape measurements. Automated imaging of cells on pretensioned microarrays using the focal planes predicted by the model produced high quality fluorescence images of cells, enabling accurate cell area quantification (<4% error) at increased speed (13×) relative to conventional methods. Our microfabrication method and simplified, linear modeling approach is readily applicable to control the deformation of similar membranes in MEMs devices, sensors, and microfluidics.

SUBMITTER: DiSalvo M

PROVIDER: S-EPMC5912951 | biostudies-literature | 2018 Apr

REPOSITORIES: biostudies-literature

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Characterization of Tensioned PDMS Membranes for Imaging Cytometry on Microraft Arrays.

DiSalvo Matthew M Harris Daniel M DM Kantesaria Saurin S Peña Alexis N AN Allbritton-King Jules D JD Cole Jacqueline H JH Allbritton Nancy L NL

Analytical chemistry 20180313 7

Polydimethylsiloxane (PDMS) membranes can act as sensing elements, barriers, and substrates, yet the low rigidity of the elastomeric membranes can limit their practical use in devices. Microraft arrays rely on a freestanding PDMS membrane as a substrate for cell arrays used in imaging cytometry and cellular isolation. However, the underlying PDMS membrane deforms under the weight of the cell media, making automated analytical microscopy (and thus cytometry and cell isolation) challenging. Here w ...[more]

PMID: 29510027

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Project description:Protein-reinforced supported bilayer membranes (rSBMs) composed of phosphatidylcholine (PC), biotin-PE and Neutravidin were used to coat hybrid microchips composed of polydimethylsiloxane (PDMS) and glass. Since the coatings required a freshly oxidized, hydrophilic substrate, a novel method to rapidly connect reservoirs using plasma oxidation was first developed and found to support up to 5.2 N cm(-2) (1.5 N) pull-off force. rSBMs were then assembled in the oxidized hydrophilic channels. The electroosmotic mobility (mu(eo)) of rSBM-coated channels was measured over a 3 h time to evaluate the stability of the coatings for microchip electrophoresis. rSBM-coated microchips with a simple cross-design had excellent properties for microchip separations, yielding efficiencies of up to 700,000 plates m(-1) for fluorescent dyes and peptides. The separation performance of rSBM and PC-coated channels was evaluated after repeatedly drying and rehydrating the channels. The separation efficiency of fluorescein on PC-coated devices decreased by 40% after one dehydration cycle and nearly 75% after 3 cycles. In contrast for rSBM-coated devices there was no significant change in the fluorescein efficiency until the third cycle (10% decreased efficiency). rSBM-coated channels were also markedly more stable when placed in a dehydrated state during long-term storage compared to PC-coated channels, and showed reduced chip failure and no reduction in performance for up to one month of dehydrated storage. Finally, rSBM-coated devices were used to perform single-cell cytometry. Microchips that had been dehydrated, stored two weeks, and rehydrated prior to use demonstrated similar performance to newly coated devices for the separation of fluorescein and carboxyfluorescein from single cells. Thus rSBM-coated devices were rugged withstanding electric fields, prolonged storage under dehydrated conditions, and biofouling by cellular constituents while maintaining excellent separation performance.

Dataset Information

Characterization of Tensioned PDMS Membranes for Imaging Cytometry on Microraft Arrays.

Publications

Characterization of Tensioned PDMS Membranes for Imaging Cytometry on Microraft Arrays.

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