A miniaturized, disposable microbial culture chip has been fabricated by microengineering a highly porous ceramic sheet with up to one million growth compartments. This versatile culture format, with discrete compartments as small as 7 ؋ 7 m, allowed the growth of segregated microbial samples at an unprecedented density. The chip has been used for four complementary applications in microbiology. (i) As a fast viable counting system that showed a dynamic range of over 10,000, a low degree of bias, and a high culturing efficiency. (ii) In high-throughput screening, with the recovery of 1 fluorescent microcolony in 10,000. (iii) In screening for an enzyme-based, nondominant phenotype by the targeted recovery of Escherichia coli transformed with the plasmid pUC18, based on expression of the lacZ reporter gene without antibiotic-resistance selection. The ease of rapid, successive changes in the environment of the organisms on the chip, needed for detection of -galactosidase activity, highlights an advantageous feature that was also used to screen a metagenomic library for the same activity. (iv) In high-throughput screening of >200,000 isolates from Rhine water based on metabolism of a fluorogenic organophosphate compound, resulting in the recovery of 22 microcolonies with the desired phenotype. These isolates were predicted, on the basis of rRNA sequence, to include six new species. These four applications suggest that the potential for such simple, readily manufactured chips to impact microbial culture is extensive and may facilitate the full automation and multiplexing of microbial culturing, screening, counting, and selection. microdish ͉ microcolony ͉ cellular assay ͉ nanoporous aluminum oxide
The strong demand for renewable energy promotes research on novel methods and technologies for energy conversion. Microfluidic systems for energy conversion by streaming current are less known to the public, and the relatively low efficiencies previously obtained seemed to limit the further applications of such systems. Here we report a microdropletbased electrostatic generator operating by an acceleration-deceleration cycle ('ballistic' conversion), and show that this principle enables both high efficiency and compact simple design. Water is accelerated by pumping it through a micropore to form a microjet breaking up into fast-moving charged droplets. Droplet kinetic energy is converted to electrical energy when the charged droplets decelerate in the electrical field that forms between membrane and target. We demonstrate conversion efficiencies of up to 48%, a power density of 160 kW m À 2 and both high-(20 kV) and low-(500 V) voltage operation. Besides offering striking new insights, the device potentially opens up new perspectives for low-cost and robust renewable energy conversion.
In this article we describe the development of a microfluidic chip to determine the concentration of spermatozoa in semen, which is a main quality parameter for the fertility of a man. A microfluidic glass-glass chip is used, consisting of a microchannel with a planar electrode pair that allows the detection of spermatozoa passing the electrodes using electrical impedance measurements. Cells other than spermatozoa in semen also cause a change in impedance when passing the electrodes, interfering with the spermatozoa count. We demonstrate that the change in electrical impedance is related to the size of cells passing the electrodes, allowing to distinguish between spermatozoa and HL-60 cells suspended in washing medium. In the same way we are able to distinguish between polystyrene beads and spermatozoa. Thus, by adding a known concentration of polystyrene beads to a boar semen sample, the spermatozoa concentrations of seven mixtures are measured and show a good correlation with the actual concentration (R(2)-value = 0.97). To our knowledge this is the first time that the concentration of spermatozoa has been determined on chip using electrical impedance measurements without a need to know the actual flow speed. The proposed method to determine the concentration can be easily applied to other cells. The described on-chip determination of the spermatozoa concentration is a first step towards a microfluidic system for a complete quality analysis of semen.
Handling microorganisms in high throughput and their deployment into miniaturized platforms presents significant challenges. Contact printing can be used to create dense arrays of viable microorganisms. Such "living arrays", potentially with multiple identical replicates, are useful in the selection of improved industrial microorganisms, screening antimicrobials, clinical diagnostics, strain storage, and for research into microbial genetics. A high throughput method to print microorganisms at high density was devised, employing a microscope and a stamp with a massive array of PDMS pins. Viable bacteria (Lactobacillus plantarum, Esherichia coli), yeast (Candida albicans) and fungal spores (Aspergillus fumigatus) were deposited onto porous aluminium oxide (PAO) using arrays of pins with areas from 5 x 5 to 20 x 20 microm. Printing onto PAO with up to 8100 pins of 20 x 20 microm area with 3 replicates was achieved. Printing with up to 200 pins onto PAO culture chips (divided into 40 x 40 microm culture areas) allowed inoculation followed by effective segregation of microcolonies during outgrowth. Additionally, it was possible to print mixtures of C. albicans and spores of A. fumigatus with a degree of selectivity by capture onto a chemically modified PAO surface. High resolution printing of microorganisms within segregated compartments and on functionalized PAO surfaces has significant advantages over what is possible on semi-solid surfaces such as agar.
This paper describes the realization process of small-size electrets by using techniques generally applied in the fabrication of integrated circuits and microsensors. In the first part of the paper, attention is paid to the different electret decay mechanisms and their relative contribution to the overall stability of miniaturized electrets. Then, a process is described by which polymer electrets such as Teflon-FEP and PTFE can be deposited and shaped in a predefined pattern on a silicon wafer. In the third part, results on the application of new materials, especially silicon dioxide (S i o s) , for use in electret applications, are presented. It appears that after an appropriate treatment of the oxide surface, its charge-stability is at least equal to that of polymer electrets such as Teflon-FEP and PTFE.
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