We present a droplet-based microfluidics protocol for high-throughput analysis and sorting of single cells. compartmentalization of single cells in droplets enables the analysis of proteins released from or secreted by cells, thereby overcoming one of the major limitations of traditional flow cytometry and fluorescence-activated cell sorting. as an example of this approach, we detail a binding assay for detecting antibodies secreted from single mouse hybridoma cells. secreted antibodies are detected after only 15 min by co-compartmentalizing single mouse hybridoma cells, a fluorescent probe and single beads coated with anti-mouse IgG antibodies in 50-pl droplets. the beads capture the secreted antibodies and, when the captured antibodies bind to the probe, the fluorescence becomes localized on the beads, generating a clearly distinguishable fluorescence signal that enables droplet sorting at ~200 Hz as well as cell enrichment. the microfluidic system described is easily adapted for screening other intracellular, cell-surface or secreted proteins and for quantifying catalytic or regulatory activities. In order to screen ~1 million cells, the microfluidic operations require 2–6 h; the entire process, including preparation of microfluidic devices and mammalian cells, requires 5–7 d.

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Di-(2-ethylhexyl)phthalate (DEHP) and total phthalate ester plasticizer levels were determined in milk, cream, butter and cheese samples from a variety of sources from three European countries (UK, Norway and Spain). Samples of milk (from Norway) obtained at various stages during collection, transportation and packaging operations showed no apparent trends in phthalate contamination with total phthalate levels (expressed as DEHP equivalents) in the raw milk of between 0.12 and 0.28 mg/kg. On processing the DEHP was concentrated in the cream at levels up to 1.93 mg/kg, whereas low fat milk contained from < 0.01 to 0.07 mg/kg. Retail dairy products (from Spain) were contaminated with < 0.01-0.55 mg/kg DEHP with a maximum total phthalate level of 3.0 mg/kg in cream samples. UK pooled milk samples from doorstep delivery (obtained from different regions of the country) contained low levels of DEHP (< 0.01-0.09 mg/kg) and total phthalate (0.06-0.32 mg/kg). Retail UK samples of cheese, butter and other fatty products varied considerably in their levels of contamination, the highest being cheese samples containing 17 mg/kg of DEHP and 114 mg/kg total phthalate. However, the majority of samples contained 0.6-3.0 mg/kg DEHP and 4-20 mg/kg total phthalate. UK cream samples contained levels of 0.2-2.7 mg/kg DEHP and 1.8-19.0 mg/kg total phthalate. The level found in these products was too high to have resulted solely from milk by concentration in the fat phase and must therefore have arisen in other ways.

The approach to assess exposure to ochratoxin A from the diet by the analysis of human plasma and urine samples has been developed. Composite duplicate diet samples from 50 individuals and corresponding plasma and urine samples were obtained over 30 days. Samples were analysed using sensitive methods capable of measuring ochratoxin A at 0.001 ng g(-1) in food, 0.1 ng ml(-1) in plasma and 0.01 ng ml(-1) in urine. Analysis of the foods indicated ochratoxin A levels contributing to an average intake in the range 0.26-3.54 ng kg(-1) bw day(-1) over the 30 days. Ochratoxin A was found in all plasma samples and in 46 urine samples. The correlation between the plasma ochratoxin A levels and ochratoxin A consumption was not significant (95% confidence limit). However, a significant correlation was found between ochratoxin A consumption and the urine ochratoxin A concentration expressed as the total amount excreted. This new work offers the possibility of using ochratoxin A in urine as a simple and reliable biomarker to estimate exposure to this mycotoxin.

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