Institute of Fundamental Technological Research

Blood is the richest source of diagnostic information. The growing interest in point-of-care analytics prompted several attempts to extract plasma from whole blood in simple diagnostic devices. The simplest method of separation is sedimentation. Here we show the first microfluidic system that uses sedimentation to extract plasma from undiluted blood and integrates execution of liquid assays on the extracted material. We present a microfluidic chip that accepts a small sample (27 μL) of whole blood, separates up to 6 μL of plasma, and uses metered volumes of plasma and of reagent (2-chloro-4-nitrophenyl-α-maltotrioside, CNP-G3) for a liquid enzymatic assay. With a custom designed channel, the system separates blood by sedimentation within few minutes of accepting the sample, mixes it with the reagent, and quantifies spectrophotometrically the product of the enzymatic reaction. As a model demonstration, we show a quantitative enzymatic α-amylase assay that is routinely used in diagnosis of pancreas diseases. The paper reports the design and characterization of the microfluidic device and the results of tests on clinically collected blood samples. The results obtained with the microfluidic system compare well to a reference bench-top analyzer.

We demonstrate a strategy for construction of high-throughput microfluidic systems generating gradations of chemistry in micro-droplets. The productivity of the systems that we propose is limited only by the maximum rate of the droplet formation, and does not need to be limited by the rate of mixing. Multilayer polycarbonate chips transform two miscible input streams A and B into N streams of droplets, containing mixtures [A]i, [B]i. Exemplary devices generate linear ([B]i ∝ i) and logarithmic gradations (ln[B]i ∝ i). We also analyze the use of the same strategy for the generation of concentration gradation in the streams of droplets comprising mixtures of liquids of different viscosities. The devices preserve the required distribution of compositions, while allowing the volume of the droplets to be tuned over almost two orders of magnitude (i.e. between 3 and 80 nL).

Microfluidics, Generation of gradients, Droplet, Viscosity