The use of two-phase flow in lab-on-chip devices, where chemical and biological reagents are enclosed within plugs separated from each other by an immiscible fluid, offers significant advantages for the development of devices with high throughput of individual heterogeneous samples. Lab-on-chip devices designed to perform the polymerase chain reaction (PCR) are a prime example of such developments. The internal circulation within the plugs used to transport the reagents affects the efficiency of the chemical reaction within the plug, due to the degree of mixing induced on the reagents by the flow regime. It has been hypothesised in the literature that all plug flows produce internal circulation. This work demonstrates experimentally that this is false. The particle image velocimetry (PIV) technique offers a powerful nonintrusive tool to study such flow fields. This paper presents micro-PIV experiments carried out to study the internal circulation of aqueous plugs in two phase flow within 762 lm internal diameter FEP Teflon tubing with FC-40 as the segmenting fluid. Experiments have been performed and the results are presented for plugs ranging in length from 1 to 13 mm with a bulk mean flow velocity ranging from 0.3 to 50 mm/s. The results demonstrate for the first time that circulation within the plugs is not always present and requires fluidic design considerations to ensure their generation.Keywords Two-phase flow Á Plug flow Á Internal circulation Á lPIV List of symbols m magnification NA numerical aperture R radius of tubing (mm) V velocity (mm/s) V avg average velocity of plug (mm/s) V max maximum velocity (mm/s) V mean bulk bulk mean velocity (mm/s) X distance (mm) Y distance (mm) d e effective diameter of particle on CCD (lm) d p diameter of particle (lm) d s point spread function (lm) n refractive index r radial position (mm)Greek symbols dx measurement uncertainty (lm) dz m measurement depth (lm) h light collection angle (rad) k 0 wavelength of light in a vacuum (nm)

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The issue of contamination of micro channel surfaces by bio fluids is a significant impediment to the development of many biomedical devices. A solution to this problem is the use of a carrier fluid, which segments the bio fluid and forms a thin film between the bio fluid and the channel wall. A number of issues need to be addressed for the successful implementation of such a solution. Amongst these is the prediction of the thickness of the film of carrier fluid which forms between the bio sample and the channel wall. The Bretherton and Taylor laws relate the capillary number to the thickness of this film. This paper investigates the validity of these laws through an extensive experimental program in which a number of potential carrier fluids were used to segment aqueous droplets over a range of flow rates. The aqueous plugs were imaged using a high speed camera and their velocities were measured. Film thicknesses were calculated from the ratio of the velocity of the carrier fluid to the velocity of the aqueous plug. The paper concludes that significant discrepancies exist between measured film thicknesses and those predicted by the Bretherton and Taylor laws.

This paper evaluates the compatibility of segmenting fluids for two phase flow applications in biomedical microdevices. The evaluated fluids are chosen due to the variations in fluid properties and cost, while also reflecting their use in the recent literature. These segmenting fluids are examined to determine their compatibility with the Polymerase Chain Reaction (PCR), through controlled experiments. The results are the first to provide a quantitative measure of segmenting fluid compatibility with PCR.

The two-phase segmented flow approach to the processing and quantitative analysis of biological samples in microdevices offers significant advantages over the single-phase continuous flow methodology. Despite this, little is known about the compatibility of samples and reactants with segmenting fluids, although a number of investigators have reported reduced yield and inhibition of enzymatic reactions depending on the segmenting fluid employed. The current study addresses the compatibility of various segmenting fluids with real time quantitative PCR to understand the physicochemical requirements of this important reaction in biotechnology. The results demonstrate that creating a static segmenting fluid/PCR mix interface has a negligible impact on the reaction efficiency, crossing threshold and end fluorescence levels using a variety of segmenting fluids. The implication is then that the previously reported inhibitory effects are the result of the dynamic motion between the segmenting fluid and the sample in continuously flowing systems. The results presented here are a first step towards understanding the limitations of the segmented flow methodology, which are necessary to bring this approach into mainstream use.