Probes quantification

Photoluminescence finds its greatest strengths as a qualitative and semiquantitative probe. Quantification based on absolute or relative intensities is difficult, although it is useful in applications where the sample and optical configurations may be carefully controlled. The necessary conditions are most easily met for analytical applica-... 

Equipment required Limited size of probe Quantification of fat/water content requires calibration... 

Bastin GF, Heijligers HIM (1991) Quantitative electron probe microanalysis of ultra-light elements (boron-oxygen). In Electron Probe Quantification. KFJ Heinrich, Newbury DE (eds) Plenum Press, New York, p 145-151... 

Deteaion limits Quantification Depth probed Lateral resolution Sample requirements... 

Detection limit Quantification Dynamic range Probing depth Lateral resolution Mass range... 

Destructive Chemical bonding Quantification Accuracy Detection limits Depth probed Depth resolution Lateral resolution Imaging/ mapping... 

Destructive Quantification Sensitivity Depth probed Lateral resolution... 

The limitations of SIMS - some inherent in secondary ion formation, some because of the physics of ion beams, and some because of the nature of sputtering - have been mentioned in Sect. 3.1. Sputtering produces predominantly neutral atoms for most of the elements in the periodic table the typical secondary ion yield is between 10 and 10 . This leads to a serious sensitivity limitation when extremely small volumes must be probed, or when high lateral and depth resolution analyses are needed. Another problem arises because the secondary ion yield can vary by many orders of magnitude as a function of surface contamination and matrix composition this hampers quantification. Quantification can also be hampered by interferences from molecules, molecular fragments, and isotopes of other elements with the same mass as the analyte. Very high mass-resolution can reject such interferences but only at the expense of detection sensitivity. 

Surface analysis by non-resonant (NR-) laser-SNMS [3.102-3.106] has been used to improve ionization efficiency while retaining the advantages of probing the neutral component. In NR-laser-SNMS, an intense laser beam is used to ionize, non-selec-tively, all atoms and molecules within the volume intersected by the laser beam (Eig. 3.40b). With sufficient laser power density it is possible to saturate the ionization process. Eor NR-laser-SNMS adequate power densities are typically achieved in a small volume only at the focus of the laser beam. This limits sensitivity and leads to problems with quantification, because of the differences between the effective ionization volumes of different elements. The non-resonant post-ionization technique provides rapid, multi-element, and molecular survey measurements with significantly improved ionization efficiency over SIMS, although it still suffers from isoba-ric interferences. 

Technique Primary probe Elemental range Type of information Depth of information Lateral resolution Sensitivity (at. %) Ease of quantification Insulator analysis Destructive UHV environment... 

A diagnostic method using fluorescence labeled DNA probes to detect and quantify the number complementary chromosomal sequences on a cellular resolution. A related technique that also allows assessment of gene amplifications, but without precise quantification of copy numbers is the chromogenic in situ hybridization (CISH). Here, instead of a fluorescent dye an enzyme that can generate a colored precipitate in the tissue samples is coupled to the DNA probe. 

The nano-scale structures in polymer layered-silicate nano-composites can be thoroughly characterized by X-ray diffraction (XRD) and transmission electron microscopy (TEM). XRD is used to identify intercalated structures. XRD allows quantification of changes in layer spacing and the most commonly used to probe the nano-composite structure and... 

D. Redecker, T. Batinic, I. S. Feder, K. Koseh, U. Schulz, P. Vinuesa, and D. Werner, Biocontrol strain Pseudomomis fluorescens W34 specific detection and quantification in the rhizo.sphere of Cucumis salivus with a DNA probe and characterization by DNA fingerprinting. 2. Natmforsch. 54c 359 (1999). 

Plasticiser/oil in rubber is usually determined by solvent extraction (ISO 1407) and FTIR identification [57] TGA can usually provide good quantifications of plasticiser contents. Antidegradants in rubber compounds may be determined by HS-GC-MS for volatile species (e.g. BHT, IPPD), but usually solvent extraction is required, followed by GC-MS, HPLC, UV or DP-MS analysis. Since cross-linked rubbers are insoluble, more complex extraction procedures must be carried out. The determination of antioxidants in rubbers by means of HPLC and TLC has been reviewed [58], The TLC technique for antidegradants in rubbers is described in ASTM D 3156 and ISO 4645.2 (1984). Direct probe EIMS was also used to analyse antioxidants (hindered phenols and aromatic amines) in rubber extracts [59]. ISO 11089 (1997) deals with the determination of /V-phenyl-/9-naphthylamine and poly-2,2,4-trimethyl-1,2-dihydroquinoline (TMDQ) as well as other generic types of antiozonants such as IV-alkyl-AL-phenyl-p-phenylenediamines (e.g. IPPD and 6PPD) and A-aryl-AL-aryl-p-phenylenediamines (e.g. DPPD), by means of HPLC. 

A prototype bDNA assay was developed for quantification of HGV/GBV-C RNA in serum (Pessoa et al, 1997). The assay employed target probes based on the relatively conserved sequence in the 5 untranslated region of the HGV/GB V-C genome. Preamplifier molecules and incorporation of isoC and isoG into the sequences common to bDNA assays were used to enhance the analytical sensitivity. The provisional limit of detection was 32,500 genome equivalents/ml based on dilutions of a 700-nucleotide synthetic HGV/GBV-C RNA transcript. The run-to-run variance of the assay was <15%. 

Finally, it should be kept in mind that quantification is often problematic in surface analysis and characterization. Firstly because some techniques are not really suited for quantification, but also in cases such as infrared spectroscopy where one does not really know precisely how deep into the material one is probing. Although, there are many good examples of semi-quantitative applications that involve measuring relative band intensities that relate to changes in a surface property. However, for problem solving revealing qualitative differences is often sufficient information to be able to identify cause and move on to look for a potential solution. 

Fig. 6.12. Fluorescence spectra of an ideal proteinase probe before (black line) and after (gray line) enzymatic cleavage. Quantification of the hydrolysis rate is performed by measuring the change of ratio in donor/acceptor emission... 

Figure 2 quantification of occluded and externally accessible mesoporosity using different probe molecules (A bulkier than B). 

Fattinger C., Roller H., Schlatter D., Wehrly P., The difference interferometer a highly sensitive optical probe for quantification of molecular surface concentration, Biosens. Bioelectron. 1993 8 99-107. 

The chemical modification of nucleic acids at specific sites within individual nucleotides or within oligonucleotides allows various labels to be incorporated into DNA or RNA probes. This labeling process can produce conjugates having sensitive detection properties for the localization or quantification of oligo binding to a complementary strand using hybridization assays. 

Fluorescently labeled DNA probes can be used for detection, localization, or quantification of target DNA sequences. In situ hybridization mapping of genomic DNA sequences can be... 

PFOA observations To evaluate MPI-MCTM model results observational data of PFOA from ship cruises in the Atlantic, Indian and Pacific Oceans were taken from literature (summarised in Yamashita et al (2008)). The data was collected between 2002 and 2006 in a global ocean monitoring initiative. Samples were taken from ocean surface water. Vertical profiles were sampled in the Labrador sea, the Mid Atlantic ocean, the South Pacific ocean and the Japanese sea, where water probes were done at several depths down to 5500 m. The limit of quantification for PFOA was determined as 6 pg/L. 

---