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BASIC SKILLS: DILUTIONS, MICROPIPETTES AND SPECTROPHOTOMETRY. Serial dilutions A sequential set of dilutions in which the stock for each dilution in the series is. • The problem we see with this method is that there is no convenient way to measure 10-3 µl accurately.

This paper reports a reconfigurable microfluidic dilution device for high-throughput quantitative assays, which can easily produce discrete logarithmic/binary concentration profiles ranging from 1 to 100-fold dilution in parallel from a fixed sample volume (e.g., 10 μL) without any assistance of continuous fluidic pump or robotic automation. The integrated dilution generation chip consists of switchable distribution and collection channels, metering reservoirs, reaction chambers, and pressure-activatable Laplace valves.

Following the sequential loading of a sample, a diluent, and a detection reagent into the individual metering chambers, the top microfluidic layer can be reconfigured to collect the metered chemicals into the reaction chambers in parallel, where detection will be conducted. To facilitate mixing and reaction in the micro-chambers, two acoustic microstreaming actuation mechanisms have been investigated for easy integratability and accessibility. Furthermore, the microfluidic dilution generator has been characterized by both colorimetric and fluorescent means. A further demonstration of the generic usage of the quantitative dilution chip has utilized a commonly available bicinchoninic acid (BCA) assay to analyse protein concentrations of human tissue extracts.

In brief, the microfluidic dilution generator offers a high-throughput high-efficiency quantitative analytical alternative to the conventional quantitative assay platforms, by simple manipulation of a minute amount of chemicals in a compact microfluidic device with a minimal equipment requirement, which can serve as a facile tool for biochemical and biological analyses in regular laboratories, point-of-care and low-resource environments. Introduction As a critical preparatory step, dilution generation has been a routine practice and gold standard to form quantitative readings of a wide variety of biological and biochemical assays. It is frequently utilized in sample preparation where samples are diluted into a series of concentrations for quantification with an assay. For instance, in a serology laboratory, patient’s serum is typically diluted 5-10 times in series to determine the exact amount of total proteins, cytokines,, creatinine, or certain antibodies present in the sample. Moreover, in the calibration step of quantitative assaying, dilution generation has been used to generate a standard curve whenever an indicator-labelled detection method is conducted. – Specifically, in the cytokine immunoassay, a standard curve can be plotted by reading fluorescence intensities of the pre-prepared serial dilutions from a known concentration, while the measurement result from the cytokine-containing sample with an unknown concentration can be interpolated from the standard curve.