Fluorescence microscopy, analytical method Applications

The most frequently applied analytical methods used for characterizing bulk and layered systems (wafers and layers for microelectronics see the example in the schematic on the right-hand side) are summarized in Figure 9.4. Besides mass spectrometric techniques there are a multitude of alternative powerful analytical techniques for characterizing such multi-layered systems. The analytical methods used for determining trace and ultratrace elements in, for example, high purity materials for microelectronic applications include AAS (atomic absorption spectrometry), XRF (X-ray fluorescence analysis), ICP-OES (optical emission spectroscopy with inductively coupled plasma), NAA (neutron activation analysis) and others. For the characterization of layered systems or for the determination of surface contamination, XPS (X-ray photon electron spectroscopy), SEM-EDX (secondary electron microscopy combined with energy disperse X-ray analysis) and... 

Electron probe and X-ray fluorescence methods of analysis are used for rather different but complementary purposes. The ability to provide an elemental spot analysis is the important characteristic of electron probe methods, which thus find use in analytical problems where the composition of the specimen changes over short distances. The examination of the distribution of heavy metals within the cellular structure of biological specimens, the distribution of metal crystallites on the surface of heterogeneous catalysts, or the differences in composition in the region of surface irregularities and faults in alloys, are all important examples of this application. Figure 8.45 illustrates the analysis of parts of a biological cell just 1 pm apart. Combination of electron probe analysis with electron microscopy enables visual examination to be used to identify the areas of interest prior to the analytical measurement. 

The results herein demonstrated the successful application of parallel combinatorial methods to generate libraries of sensing SAMs covalently immobilized in the wells of a glass microtiter-plate. The fluorescence pattern after exposure of the array to different metal ion solutions allows identification of Cu2+, Co2+, Ca2+, Zn2+ and Pb2+ at 10"4 M concentration by laser confocal microscopy and fluorescence laser scanner. The collection of the unselective response of the monolayers in the presence of the cations generates a characteristic fluorescent pattern, a fingerprint of each analyte in the array. 

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