Detection of membrane-bound tumor necrosis factor (TNF): an analysis of TNF-specific reagents.

Tumor necrosis factor (TNF) exists in two bioactive forms, the membrane integrated form and the proteolytically derived soluble cytokine. Both forms of TNF are involved in a variety of different physiological and pathophysiological situations. Here we analyzed different human and mouse TNF-specific reagents for their ability to determine the expression of membrane-expressed TNF. The data prove some antibodies to be very useful for the analysis of transmembrane TNF expression because these antibodies distinguish between the transmembrane form of TNF and soluble TNF bound to cellular TNF receptors. In addition, we found that recombinant human TNF receptor fusion proteins are advantageous tools to analyze both human and mouse transmembrane TNF expression.

Optical oxygen sensor instrumentation based on the detection of luminescence lifetime.

Optical oxygen sensors are mainly based on the principle of luminescence quenching. In contrast to arready existing intensity-based systems, the measurement of the luminescence lifetime provides certain advantages, such as insensitivity to photobleaching or leaching of the dye, or changes in the intensity of excitation light. This facilitates the use of simple optical systems or optical fibres. A new family of oxygen-sensitive dyes, the porphyrin-ketones, has been introduced, which exhibits favorable spectral properties and decay times in the order of tens and hundreds of microseconds. This allows the use of simple optoelectronic circuitry and low-cost processing electronics. An optical oxygen sensor module has been developed with the dimensions of only 120 x 60 x 30 mm. The prototype is based on the measurement of the decay time of the luminophore by measuring the phase shift between the square-wave excitation and the detected square-wave of the emission coming from the sensor. The instrument is based on semiconductor devices (light-emitting diodes, photodiodes) and may be used for the detection of oxygen in gaseous or liquid samples. The measurement range of the device is from 0 to 200 hPa oxygen partial pressure with a resolution of < 1 hPa over the whole measurement range. The overall measurement accuracy of < +/- 1 hPa has been obtained for periods of 24 h of continuous measurement in a thermostatted environment. The sensor response times t90 are typically < 1 s for gases and 0.5 to 5 min for liquid samples.