A multiplexed immunoassay system based upon reciprocating centrifugal microfluidics

Abstract

A novel, centrifugal disk-based micro-total analysis system (μTAS) for low cost and high throughput semi-automated immunoassay processing was developed. A key innovation in the disposable immunoassay disk design is in a fluidic structure that enables very efficient micro-mixing based on a reciprocating mechanism in which centrifugal acceleration acting upon a liquid element first generates and stores pneumatic energy that is then released by a reduction of the centrifugal acceleration, resulting in a reversal of direction of flow of the liquid. Through an alternating sequence of high and low centrifugal acceleration, the system reciprocates the flow of liquid within the disk to maximize incubation/hybridization efficiency between antibodies and antigen macromolecules during the incubation/hybridization stage of the assay. The described reciprocating mechanism results in a reduction in processing time and reagent consumption by one order of magnitude.

Biosensing enhancement of dengue virus using microballoon mixers on centrifugal microfluidic platforms

“…space on the disc as an air chamber that it used to generate pneumatic energy must be accommodated. Nevertheless, by means of the same air chamber the mixer could pump the mixed liquids against the centrifugal force to other reservoirs and therefore enable more complex bioassays on the disk (Noroozi et al, 2009(Noroozi et al, , 2011. The mixing techniques on centrifugal microfluidics discussed here are useful approaches that have been employed, for example, in quantifying nitrate and nitrite levels in blood, reparatory virus detection, detection of the antibiotic resistance gene mecA of Staphylococcus aureus and several enzyme linked immunosorbent assays (ELISA) Kim et al, 2013;Kinahan et al, 2014;Lee et al, 2011;Lutz et al, 2010;Noroozi et al, 2009Noroozi et al, , 2011Nwankire et al, 2013Nwankire et al, , 2014Siegrist et al, 2010Siegrist et al, , 2006.…”

Theory, fabrication and applications of microfluidic and nanofluidic biosensors

“…As an example, figure 1 [41] shows a biosensor that can be incorporated in a wristwatch and a cassette-type biosensor with multiple unit operations on a single platform. Other platforms such as those modelled after compact discs (CDs) that use centrifugal forces have also been developed [65]. This integration of multiple unit operations on a single platform is the hallmark of biosensors as these are typically considered multi-functional, multi-component devices [38,66].…”

Lab-on-a-CD: A Fully Integrated Molecular Diagnostic System

“…In 2011, Noroozi et al was able to integrate this technology into a multiplexed LoD immunoassay to improve Burkholderia detection, illustrating the impact this mechanism can have on sample preparation. 35 Furthermore, Aeinehvand et al incorporated a microballoon for flow reciprocation. 87 The microballoon mixer required a smaller disc footprint than the reciprocating flow mixer by Noroozi et al and reduced mixing time from 170 min of diffusion-based mixing to less than 23 s.…”

“…Capability to effectively store both lyophilized and liquid reagents on the same disc. , siphon, 32 micropulley, 33 and pneumatic [34][35][36] Wax, 42 thermo-pneumatic, 45 electrolysis; 46 magnetic, and laser 47 Dissolvable films, 52 paper siphon, 61 and check valves 43,63 Each type of valve has trade-offs regarding complexity, cost, and reliability.…”

“…Reciprocating the sample volume also introduces chaotic advection, which reduces the reaction time as compared to passive diffusion. 35 Pneumatic pumping was further improved with the implementation of the latex microballoon pump introduced by Aeinehvand et al (see Fig. 5).…”