Abstract
We report a device to fill an array of small chemical reaction chambers (microreactors) with reagent and then seal them using pressurized viscous liquid acting through a flexible membrane. The device enables multiple, independent chemical reactions involving free floating intermediate molecules without interference from neighboring reactions or external environments. The device is validated by protein expressed in situ directly from DNA in a microarray of ~10,000 spots with no diffusion during three hours incubation. Using the device to probe for an autoantibody cancer biomarker in blood serum sample gave five times higher signal to background ratio compared to standard protein microarray expressed on a flat microscope slide. Physical design principles to effectively fill the array of microreactors with reagent and experimental results of alternate methods for sealing the microreactors are presented.
Highlights
We report a device to fill an array of small chemical reaction chambers with reagent and seal them using pressurized viscous liquid acting through a flexible membrane
Using the device to probe for an autoantibody cancer biomarker in blood serum sample gave five times higher signal to background ratio compared to standard protein microarray expressed on a flat microscope slide
The microreactor array platform consists of an array of functionalized microreactors in a microscope slide format and a device for filling the microreactors with reagent and sealing them
Summary
We report a device to fill an array of small chemical reaction chambers (microreactors) with reagent and seal them using pressurized viscous liquid acting through a flexible membrane. Around a million experiments are performed simultaneously for gene expression analysis[2] and around a billion for generation DNA sequencing[3] These high throughput experiments are based on molecules tethered to a surface. Many different biochemical reactions occur simultaneously depending on cell type, cell cycle or external stimuli Unravelling this complexity and its effect on human health requires high throughput experimental platforms that can simultaneously study thousands of biochemical reactions involving untethered, free floating, molecular compounds. Proteins can be expressed outside of living cells by subjecting gene DNA to ‘cell-free’ in vitro coupled transcription and translation (IVTT) reagent. The device is amenable to production scale processing of microreactor array slides
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