Temperature change rate actuated bubble mixing for homogeneous rehydration of dry pre-stored reagents in centrifugal microfluidics. Issue 2 (3rd January 2018)

• Record Type: Journal Article
• Authors: Hin, S.; Paust, N.; Keller, M.; Rombach, M.; Strohmeier, O.; Zengerle, R.; Mitsakakis, K.
• Is Part Of: Lab on a chip. Volume 18:Issue 2(2018)
• Page Start: 362
• Page End: 370
• Publication Date: 2018-01-03
• Subjects: Miniature electronic equipment -- Periodicals; Combinatorial chemistry -- Periodicals; Biotechnology -- Periodicals; 543.0813
• DOI: 10.1039/c7lc01249g
• Languages: English
• ISSNs: 1473-0197

Abstract

Abstract : Temperature change rate as actuation principle for a bubble mixer in centrifugal microfluidics minimizes external means required. We applied the new bubble mixer to the rehydration of dry reagents for nucleic acid amplification. Abstract : In centrifugal microfluidics, dead volumes in valves downstream of mixing chambers can hardly be avoided. These dead volumes are excluded from mixing processes and hence cause a concentration gradient. Here we present a new bubble mixing concept which avoids such dead volumes. The mixing concept employs heating to create a temperature change rate (TCR) induced overpressure in the air volume downstream of mixing chambers. The main feature is an air vent with a high fluidic resistance, representing a low pass filter with respect to pressure changes. Fast temperature increase causes rapid pressure increase in downstream structures pushing the liquid from downstream channels into the mixing chamber. As air further penetrates into the mixing chamber, bubbles form, ascend due to buoyancy and mix the liquid. Slow temperature/pressure changes equilibrate through the high fluidic resistance air vent enabling sequential heating/cooling cycles to repeat the mixing process. After mixing, a complete transfer of the reaction volume into the downstream fluidic structure is possible by a rapid cooling step triggering TCR actuated valving. The new mixing concept is applied to rehydrate reagents for loop-mediated isothermal amplification (LAMP). After mixing, the reaction mix is aliquoted into several reaction chambers for geometric multiplexing. As a measure for mixing efficiency, the mean coefficient of variation (C̄V̄, n = 4 LabDisks ) of the time to positivity ( t p ) of the LAMP reactions ( n = 11 replicates per LabDisk ) is taken. The C̄V̄ of the t p is reduced from 18.5% (when using standard shake mode mixing) to 3.3% (when applying TCR actuated bubble mixing). The bubble mixer has been implemented in a monolithic fashion without the need for any additional actuation besides rotation and temperature control, which are needed anyhow for the assay workflow.