Signal detection probe

An alternative to a saddle coil would be a solenoid coil which, however, would have to be oriented perpendicularly to the magnet bore and thus to the physical axis of the probe assembly. Due to spatial constraints, such an arrangement complicates considerably sample insertion, especially when the sample temperature has to be controlled and the assembly has to include an enveloping dewar for temperature control of the sample.  

In our final realization (Fig. 18), the probes use the Helmholtz coil geometry, favoring ease of use and efficient sample temperature control over a wide range of temperature values. The tunable, broad-band probe is inserted into the magnet from below and fixed to the bottom part of the magnet assembly in a simple way reminiscent of most high-resolution NMR systems. Thanks to this design, it is possible to use standard 10 mm NMR sample tubes which are inserted comfortably from above without any need to manipulate the probe. 

By the term console we intend all the electronic functional blocks used in a traditional NMR spectrometer, excluding the field control. In particular, it includes the following subsystems  

Actually, a console to be used with an FFC NMR relaxometer does not differ much from any conventional general-purpose NMR console. With the exception of the relatively simple interfaces controlling the magnet power supply and thus the field, all other hardware units are mostly the same as in any sufficiently versatile NMR spectrometer or relaxometer (what does differ quite a lot, of course, is the application software). 

For this reason, a detailed description of the console in this paper is superfluous, except for a brief list of those features which, in our opinion, any research-grade FFC console should possess in order to guarantee maximum versatility of NMR dispersion measurements. 

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Automation of ISH from the denaturation of chromosomal DNA on slides to the detection of fluorescent signals after probe hybridization is an approach to further simplification of this technique (H4). 

Before starting experiments with human or animal tissue samples, it is extremely important to optimize in vitro experimental conditions. With a purified template nucleic acid, standardize RT and PCR conditions. Check the specificity and crossreactivity of primers and probes. Sometimes it is necessary to alter MgCl2 concentration under in situ reaction conditions. The blocking reagent for filter hybridization could be different than the in situ protocol. (I use 1 % purified casein solution for filter hybridization and 3% BSA for in situ signal detection.)... 

It is apparent that signal amplification provides increased sensitivity over direct labeling. This is especially true for fluorescent-based assays. One of the most sensitive signal detection technologies is the immunoRCA (Schweitzer et al., 2000). Rolling circle amplificahon (RCA) is combined with antibody detection. RCA involves the amplification of circularized oligonuceotide probes under isothermal conditions by DNA polymerase (Lizardi et al., 1998). With immunoRCA, the 5 primer is attached to the reporter antibody. Initiation of the amplification starts when circular DNA template binds to the attached primer. 

Common haptens used for labeling DNA probes for BISH assays are biotin, DIG, DNP, FITC, and Texas Red. Based on the size of your DNA targets, you may choose from a direct detection or an indirect detection for BISH assays. In general, an indirect detection system can provide better sensitivity compared to a direct detection system. For an indirect detection, you need to select a combination of two antibodies raised with two different animal species, such as a mouse anti-DIG antibody and a rabbit anti-DNP antibody, so that enzyme-labeled anti-mouse antibody and anti-rabbit antibody can be applied for signal detection. If a direct BISH detection is going to be applied, anti-hapten antibodies raised in the same animal species that are labeled with either AP or HRP enzyme molecules... 

The controversy over the degree to which radioactive probes are more sensitive has not been fully resolved. In any case, microwave pretreatment enhances ISH signal detection of RNA and DNA whether radiolabeled or nonradioactive probes are used both methods are presented later. Furthermore, a number of approaches is available to increase the sensitivity of the nonradioactive ISH procedures (for a review, see Komminoth and Werner, 1997) some of these approaches are discussed below. 

Kim NH, Baek TJ, Park HG, Seong GH (2007) Highly sensitive biomolecule detection on a quartz crystal microbalance using gold nanoparticles as signal amplification probes. Anal... 

All NMR spectra were collected on a Varian Unity 500 MHz spectrometer ( H frequency 499.8 MHz) equipped with a 5 mm inverse detection probe. Sample concentrations were typically 1 -2 mM and sample temperatures maintained at 25 C (unless otherwise noted). Sample pH was typically 3.5 - 4.0. Onedimensional H data were acquired with a H sweepwidth of 6000 Hz and an acquisition time of 2.3 seconds. The residual water signal was suppressed by presaturation. H DQF-COSY, NOESY and TOCSY (15) spectra were collected and processed using standard methods. All chemical shifts were referenced relative to internal DSS. 

Near-field scanning optical microscopy (NSOM) has been developed as a combination of scanning probe microscope and optical microscope in which the spatial resolution is determined by scanning probe microscope resolution while the signals detected are coming from several optical interactions. As a result, NSOM has achieved a higher spatial resolution than that of the classical optical microscopy that uses a conventional lens, which is strictly limited by the diffraction... 

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