Quantitative analysis autoradiography

Irons, R. D. and E. A. Gross, Standardization and calibration of whole-body autoradiography foy routine semi-quantitative analysis of the distribution of C-labelled compounds in animal tissues. Toxicol. Appl. Pharm., 1980, in press. 

For MK2, one may use a peptide substrate (see, e.g., Manke et al, 2005) or recombinant heat shock protein (hsp) 25 (Stokoe et al, 1992). For the peptide substrate, analysis is as described for the p90RSK assay. If using hsp27 as substrate, samples of the reaction products are analyzed by SDS-PAGE and autoradiography or quantitation performed by phosphorimager. 

Domer P. Photometric methods in quantitative autoradiography, in Microautoradiography and Electron Probe Analysis. Their Application to Plant Physiology (Luttge U, ed.), Springer Verlag, New York, 1972, pp. 7M8. 

Gray, S. N., Dighton, J., Olsson, S. Jennings, D. H. (1995). Real-time measurement of uptake and translocation of 137Cs within mycelium of Schizophyllum commune Fr by autoradiography followed by quantitative image-analysis. New Phytologist, 129, 449-65. 

The analysis of samples in biological matrices is extremely powerful compared with drawbacks such as narrow concentration range for quantitative measurements and long time required for spot detection. Digital autoradiography can be used in hyphenated off-line and... 

Quantitate the radioactivity, as a measure of RT activity in 1 pL of the culture supernatant, either by liquid scintillation counting (cut out the spots and place them directly into scintillation fluid), by phosphorimage analysis and quantitation of relative pixel units, or by autoradiography and laser densitometry. 

Coincident with this new technique for procurement of human bone biopsies was the development of quantitive methods of bone analysis.12 These methods include histochemical analysis of both decalcified and unde-calcified42 48 bone sections, microradiography,44 tetracycline labeling45 and autoradiography.42 The latter two techniques require administration of a tetracycline antibiotic or isotopic tracer prior to procurement of the biopsy. Undecalcified thin sections, prepared with the use of a Jung microtome after the bone core is fixed, dehydrated and embedded in methacrylate,45 are analyzed by intersect and point count methods46 47 which permit three-dimensional assessment.48 49 Tetracycline antibiotics deposit in vivo in sites of bone formation constituting markers which can be studied in undecalcified sections by fluorescence microscopy.45 47 This represents the safest and best tissue time marker for microscopic measurement of bone formation dynamics. 

For whole brain or for a brain region in which analysis is by quantitative autoradiography or PET, we have an overall k equal to... 

Is subjected to autoradiography or phosphorlmager analysis. The radioactive bands correspond to newly synthesized proteins, which have Incorporated the radiolabeled amino acid. Alternatively, the proteins can be resolved by liquid chromatography, and the radioactivity In the eluted fractions can be determined quantitatively with a counter. 

Techniques for detection and quantitation of zones are an important aspect of electrophoretic analysis. The most common approaches to detection involve the use of stains, autoradiography of radio-labelled analytes, or immunoreaction with specially prepared antisera. In the case of electrophoresis in free solution, on-line detection devices can be used. 

At the completion of 2-D PAGE, gels are fixed and the separated proteins visualized by any appropriate technique (e.g. silver staining, autoradiography). It then remains to extract qualitative and quantitative data from the resulting complex 2-D protein patterns. Simple visual inspection can provide only limited information and it is usually necessary to use sophisticated computer analysis systems (71, 106, 10 7). 

As previously mentioned, neither the incorporation of radioactive amino acids into embryo proteins nor the presence of amino acids in the immediate environment of the embryo necessarily indicate that free amino acids contribute significantly to the nutritional needs of the embryo. Nevertheless, it should be noted that Mintz (1964), employing autoradiography, observed in the cultured mouse embryo an increase in amino acid incorporation from fertilization to the blastocyst stage. This observation was essentially confirmed (Brinster, 1971) for the mouse when quantitative procedures for the analysis of radioactivity were used. During cleavage of the rabbit embryo in vitro (Manes and Daniel, 1969),... 

Sometimes, rather than removing all the adsorbent as in zonal analysis, the sample spots/zones can be removed after being located by a suitable detection method such as UV absorption, exposure to iodine vapor, spray reagents, or autoradiography (6). This method can be used in a quantitative way for the preparative isolation of individual components. 

Another major step forward for radio-TLC came in the early 1980s when the so-called linear analyzer was introduced. This instrument was easier to use and more sensitive than the old scanners and was automated to the extent that up to four plates could be run overnight. More details are given below. As a consequence, improved quantitative results were obtained and analysis time was shortened. However, resolution was still not as good as that obtained by using autoradiography, and two-dimensional plates could not be easily evaluated. 

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