Cameras and imaging

Next Generation Sequencing Device Materials

January 29, 2019

Next Generation Sequencing [NGS], is the term coined for the massively parallel sequencing resulting in gigabytes of sequence data per day to quickly identify gene sequences. NGS is being harnessed for everything from personalized medicine to rapidly identifying microbial food contaminants [1]. The best Next Generation Sequencing device materials continue to shift as increasing innovative fabrication and detection approaches to NGS emerge.

Image 1. Microscope Slide sized Generic NGS Flow Cell provided by IMT AG. Sub-micron structures are generated on a 200mm glass wafer. This wafer is bonded together with a second wafer containing isotropically etched channels, fluidic access holes and an anti-fouling coating; i.e. PEG.

Many technologies use optical methods – and in this use case, glass offers the best optical properties. NGS sequencers such as those by Illumina and Pacific Biosystems continue to use glass. Pacific Biosystems is pushing the limits of detection for its new single molecule sequencing product, so background fluorescence and scattering are critical parameters to performance. Emerging next gen sequencing powerhouse, LaserGen, utilizes fluorescence for detection with glass microfluidic flow cells.

Illumina managed to reach the $1,000 genome target by harnessing the power of glass fabrication. By structuring the glass surface, creating patterned flow cells, it was possible for Illumina to increase the density of the sequencing space and reduce image acquisition time since patterned images are easier to overlap during data acquisition rather than unstructured images [2]. Important features to this success are the high accuracy of sub-micron patterning on a wafer level. This enables the extremely dense packaging of the sequencing space within the microfluidic chip, reaching the optical resolution limit of conventional fluorescent imaging systems. Details of the patterned flow cells at Illumina, Inc. can be found on the Illumina website [3].

IMT AG’s glass wafer processing capabilities allows an automated and scalable production of complex, multilayer microfluidic flow cells for life sciences applications including protein, DNA and cell handling and analysis.

Important features are the high accuracy of sub-micron patterning on a wafer level that enables the extremely dense packaging of the sequencing space within the microfluidic chip, reaching the optical resolution limit of conventional fluorescent imaging systems.

The capabilities of sub-micron structuring within IMT is well established and documented from our classical business, i.e. optical components. This technology is based on internal IMT processes demonstrates an excellent and flexible production of such structures on an industrial scale but with the advantage of utilizing non-MEMS materials, e.g. Glass, Gold, Silanes or other immobilization chemistry.

The path companies chose to take, due to lack of a one-stop-shopping proposal in the market, is to define and utilise a rather intricate and fragmented in-sourcing production of flow-cell sub-components and doing the functionalization and sealing of the flow cells in house. The latter is chosen mainly out of two reasons:

  • The companies prefer not to give the specific binding chemistry to vendors in fear of losing competitive advantage
  • Non available vendors with such proposal.

Image 2: Rendered image of a 2-level NGS flow-cell illustrating the possiblel complete workflow provided by IMT AG.

IMT AGs Proposal to the Life Science and Diagnostics market:

NGS flow cells must be manufactured to custom dimensions, lay-out, and bio-functionalized pattern, with micrometer or sub-micrometer feature sizes, and sealed without disrupting the bio-functionalization.

IMT AG considers it as a unique selling proposal into the Life Science and Diagnostics market performing the complete workflow in house at IMT AG, freeing Life Science and Diagnostics companies from developing complex in-house production steps. We offer complete flow cells, customizable to customer’s dimensions, lay-out and bio-functionalized pattern, in micrometer or in sub-micrometer size, in sealed flow cells. The bio-functionalized molecule can be a say amino-silane alone or on top of a dielectric or metallic coating. The rest of the flow cell channel network is coated by an anti-fouling coating, say a PEG. Each customer can define her specific binding chemistry for her specific analyte; turning the generic platform to a customer platform.