Mid-IR synchrotron radiation for molecular specific detection in microchip-based analysis systems.

Microstructures constructed from SU-8 polymer and produced on CaF(2) base plates have been developed for microchip-based analysis systems used to perform FTIR spectroscopic detection using mid-IR synchrotron radiation. The high brilliance of the synchrotron source enables measurements at spot sizes at the diffraction limit of mid-IR radiation. This corresponds to a spatial resolution of a few micrometers (5-20 microm). These small measurement spots are useful for lab-on-a-chip devices, since their sizes are comparable to those of the structures usually used in these devices. Two different types of microchips are introduced here. The first chip was designed for time-resolved FTIR investigations of chemical reactions in solution. The second chip was designed for chip-based electrophoresis with IR detection on-chip. The results obtained prove the operational functionality of these chips, and indicate the potential of these new devices for further applications in (bio)analytical chemistry.

Design, simulation and application of a new micromixing device for time resolved infrared spectroscopy of chemical reactions in solution.

We present a novel micromachined fast diffusion based mixing unit for the study of rapid chemical reactions in solution with stopped-flow time resolved Fourier transform infrared spectroscopy (TR-FTIR). The presented approach is based on a chip for achieving lamination of two liquid sheets of 10 microm thickness and approximately 1 mm width on top of each other and operation in the stopped-flow mode. The microstructure is made on infrared transmitting calcium fluoride discs and built up with two epoxy negative photoresist layers and one silver layer in between. Due to the highly laminar flow conditions and the short residence time in the mixer there is hardly any mixing when the two liquid streamlines pass through the mixing unit, which allows one to record a mid-IR transmission spectrum of the analytes prior to reaction. When the flow is stopped, the reactant streams are arrested in the flow-cell and rapidly mixed by diffusion due to the reduced interstream distances and the reaction can be directly followed with hardly any dead time. On the basis of two model reactions-neutralisation of acetic acid with sodium hydroxide as well as saponification of methyl monochloroacetate-the performance of the mixing device was tested revealing proper functioning of the device with a time for complete mixing of less than 100 ms. The experimental results were supported by numerical simulations using computational fluid dynamics (CFD), which allowed a reliable, quantitative analysis of concentration, pressure and flow profiles in the course of the mixing process.

[Miniature thin layer biosensors which are sensitive to glutamate and glutamine]. / Miniatiurnye tonkoplenochnye biosensory, chuvstvitel'nye k glutaminovoi kislote i glutaminu.

Integrated thin film biosensors for simultaneous measurement of L-glutamine and L-glutamate in a micro-flow cell were developed. Due to a novel glutaminase with an activity optimum in the neutral pH range a direct monitoring of glutamine in mammalian cell culture medium could be performed. The glutamine bienzyme sensor was prepared by coimmobilization of glutaminase with glutamate oxidase within a photo patterned poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogel membrane. The sensor response is linear in the concentration range of 55 mmol to 10 mmol glutamine/l. Additionally a glutamate biosensor is integrated on the sensor chip for difference measurement of possible glutamate interferences. The sensor chip can be used for at least 300 measurements without any alteration in the performance of its sensors. A new sensor chip with an integrated flow cell provides the possibility of simultaneous measurement of four different parameters at a cell volumina of 1 ml. Completing the microsystem a battery operated surface mounted device (SMD) potentiostat was developed to get a "lab on chip".

Thin-film microbiosensors for glucose-lactate monitoring.

A miniaturized device for simultaneous measurement of glucose and lactate levels was produced by means of photopatterning of enzyme-containing photosensitive membrane precursors. This device shows no cross-talk and a lifetime for both the glucose and the lactate sensors of more than 2 weeks when continuously operated in undiluted bovine serum. Linear response ranges of up to 40 mM for glucose and 25 mM for L-lactate, in combination with 95% response times of < 30 s, were realized. The devices are mass produced by means of thin-film technology on flexible carriers to give catheter-type multisensing devices for in vivo applications. Ex vivo experiments, performed with human volunteers, where the device was continuously operated in an extracorporeal, undiluted, heparinized blood stream for 6 h, gave a correlation of r > 0.98 with respect to laboratory techniques. Subcutaneous measurements of glucose levels in pigs were close to the corresponding blood levels obtained without in vivo calibration.

Miniaturized thin film glutamate and glutamine biosensors.

Integrated thin film biosensors were developed for the simultaneous measurement of L-glutamine and L-glutamate in a mu-flow cell. Due to a novel glutaminase with an activity optimum in the neutral pH range, direct monitoring of glutamine in a mammalian cell culture medium could be performed. The glutamine bienzyme sensor was prepared by co-immobilization of glutaminase with glutamate oxidase within a photopatterned poly(2-hydroxyethyl methacrylate) (pHEMA) hydrogel membrane. The sensor response was linear in the concentration range of 50 mumol to 10 mmol glutamine/l. Additionally, a glutamate biosensor was integrated on the sensor chip for difference measurement of possible glutamate interferences. The sensor-chip could be used for at least 300 measurements without any alteration in the performance of its sensors. A new sensor-chip with an integrated flow cell provided the possibility of simultaneous measurement of four different parameters at a cell volume of 1 microliter. In order to complete the microsystem, and in order to obtain a "lab on chip", a battery operated surface mounted device (SMD) potentiostat was developed.

Miniaturized integrated biosensors.

Miniaturized integrated thin-film biosensors were developed for use in clinical analyzers and for in vivo applications. A glucose and a lactate sensor were integrated with a pH-sensor on a flexible substrate. Both enzyme sensors are based on the electrochemical measurement of H2O2 produced by the enzymes glucoseoxidase and lactateoxidase respectively. The solid state pH-sensor uses a neutral carrier membrane. The intended application of this device is the monitoring of metabolic parameters in the intensive care unit and the operation theater and the use as a sensor module in clinical analyzers. The glucose-, lactate- and pH-sensor was tested in buffer solutions and undiluted serum showing excellent performance.