Detection probes

During the optical coat work stress examination method the upper plate of the head of some of the bolts was covered with an optical coat work (Fig. 4). On the head of some other bolts strain gauges were stuck which measured the plain biaxial stress state in the middle of the top surface of the head of the bolt (3.5 x 3 mm). The magnetic probe detected average stresses up to 0.1 mm depth in an area of 14 mm diameter in the middle of the head of the bolt. 

Range of elements Destructive Depth probed Detection limits... 

Information Element range Destructive Lateral resolution Depth profiling Depth probed Detection limits Quantitative Imaging... 

Bej, A. K. Mahbubani, M. H. DiCesare, J. L. Atlas, R. M. Polymerase chain reaction-gene probe detection of microorganisms by using filter-concentrated samples. Appl. Environ. Microbiol. 1991,57, 3529-3534. 

Figure 4.7 shows the structures of important carotenoids (all-E) lutein, (all-E) zeaxanthin, (all-E) canthaxanthin, (all-E) p-carotene, and (all-E) lycopene. Employing a self-packed C30 capillary column, the carotenoids can be separated with a solvent gradient of acetone water=80 20 (v/v) to 99 1 (v/v) and a flow rate of 5 pL min, as shown in Figure 4.8 (Putzbach et al. 2005). The more polar carotenoids (all-E) lutein, (all-E) zeaxanthin, and (all-E) canthaxanthin elute first followed by the less polar (all-E) p-carotene and the nonpolar (all-E) lycopene. Figure 4.9 shows the stopped-flow II NMR spectra of these five carotenoids. The chromatographic run was stopped when the peak maximum of the compound of interest reached the NMR probe detection volume.

The power and advantages of assessing cellular processes at their most fundamental level is propelling the science of oligonucleotide probe detection into one of the most prominent positions in bioconjugate chemistry. Oligonucleotide arrays containing hundreds or thousands of... 

Figure 27.1 Three common nucleoside triphosphate derivatives that can be incorporated into oligonucleotides by enzymatic means. The first two are biotin derivatives of pyrimidine and purine bases, respectively, that can be added to an existing DNA strand using either polymerase or terminal transferase enzymes. Modification of DNA with these nucleosides results in a probe detectable with labeled avidin or streptavidin conjugates. The third nucleoside triphosphate derivative contains an amine group that can be added to DNA using terminal transferase. The modified oligonucleotide then can be labeled with amine-reactive bioconjugation reagents to create a detectable probe.

When photobiotin is irradiated in the presence of DNA the reaction process nonselectively couples a biotin label to every 100-200 base residues. The result is an oligonucleotide probe detectable by the use of (strept)avidin conjugates. The uses of photobiotin for DNA or RNA modification are summarized in Chapter 11, Section 4. 

Mclnnes, J.L., Dalton, S., Vize, P.D., and Robins, A.J. (1987) Non-radioactive photobiotinlabeled probes detect single copy genes and low abundance mRNA. Bio/Technology 5, 269-272. 

Sumi H (1998) V-I characteristics of STM processes as a probe detecting vibronic interactions at a redox state in large molecular adsorbates such as metalloproteins. J Phys Chem B 102 1833-1844... 

Some fluorescent DNA stains can also be used for chromosome counterstaining, for detection of hybridized metaphase or interphase chromosomes in fluorescence in situ hybridization assays or for identifying apoptotic cells in cell populations (http //probes.invitrogen.com/handbook/sections/0806.html). For instance, Vybrant Apoptosis Assay Kit 4 (Molecular Probes) detects apoptosis on the basis of changes that occur in the permeability of cell membranes. This kit contains ready-to-use solutions of both YO-PRO-1 and propidium iodide nucleic acid stains. YO-PRO-1 stain selectively passes through the plasma membranes of apoptotic cells and labels them with moderate green fluorescence. Necrotic cells are stained red-fluorescent with propidium iodide. 

Various methods are used for read-out. Micelle formation and dissociation may be detected by means of a fluorescence probe detect-... 

The paramagnetic probes detected heterogeneity of swelling behavior in smectites that was not obvious from X-ray diffraction data, presumably caused by variation of charge density on different clay layers within the same smectite sample. 

Kadirvel M, Arsic B, Freeman S, Bichenkova EV (2008) Exciplex and excimer molecular probes detection of conformational flip in a myo-inositol chair. Org Biomol Chem... 

Table 3. DNA probe detection and distribution of naphthalene biodegradative bacteria in Manufactured Gas Plant soils...

Figure 19.1. Schematic diagram of a general pump-probe-detect laser spectrometer suitable for picosecond electronic absorption, infrared (IR) absorption, Raman, optical calorimetry, and dichroism measurements. For picosecond fluorescence—a pump-detect method, no probe pulse needs to be generated.

Figure 20.2. Schematic outline of typical pump-probe-detect experiments with femtosecond pulses, a molecular beam source, and mass spectrometric detection of transient species. Computer control and data processing instruments, as well as various optical components, are not shown. The time separation Af between pump and probe pulses is dictated by the difference in optical path lengths. Ad, traversed by the two components of the original pulse.

The pump-probe-detect arrangements for the femtosecond experiments was similar to those described above. When cyclobutanone was pumped with two photons of a X = 307-nm femtosecond pulse, two consecutive C—CO bond cleavages led to the formation of the trimethylene diradical, detected as an easily ionized transient at 42 amu, with buildup and decay times of 120 20 fs. The decay presumably involves isomerizations to cyclopropane and to propylene— structures not ionized by the probe pulse and thus undetected during the experiment. 

Oligonucleotide probe detects Hb S allele. [Note indicates radioactive label.]... 

In addition to biotin, a digoxigenylated derivative of dUTP was also synthesized. This derivative of dUTP can be incorporated into DNA by Pol I (or the Klenow fragment of Pol I). Therefore, digoxigenin-labeled DNA probes can be prepared by nick translation or random primed-labeling methods developed for the biotin system. It is almost certain that more nonradioactive alternatives to biotin and digoxigenin will be developed in the future. Chemiluminescent methods for nonradioactive probe detection are now widely being used... 

Corrosion probes detect and measure the amount of corrosion occurring at a given point in a system and can be used to estimate the total amount of corrosion and the type of corrosion anticipated. Probes are available for use in a wide variety of temperature and pressure conditions. 

The Rab21 rice cDNA probe detects three different transcripts. Two of them (1.5 and 0.9 kb) are cold-induced. The smallest of these transcripts corresponds in size to the Rab21 transcript described (Mundy Chua, 1988). In barley the same cDNA clone detects a cold induced transcript of 2.4 kb. 

Tsourkas et al. (2003) reported dual FRET molecular beacon assays, where the donor probe was labeled with either Eu3+ or Tb3+ complex of DTPA-csl24-ethylenediamine (and no quenchers attached). For the Eu3+ complex, the acceptor probe was Cy5-labeled (and no quenchers attached) and for the Tb complex, the acceptor probe was labeled with Cy3 or ROX as a fluorophore and with dabcyl as a quencher. They demonstrated that these pairs of probes detected DNA targets ( 50-mer) with high S/N. 

In the fluorescence microscope, careful consideration of the sample and system components is necessary to specify the correct filters for probe detection. Use of multiband dichroics and emission filters in a stationary turret with single exciters in an external slider or filter wheel can give near simultaneous probe detection with no registration shift, but there are likely compromises in overall brightness, color balance difficulty, and reduced resolution of the color CCD camera. 

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