CALIBRATION OF DENSITOMETRY IN RADIO-ISOTOPIC IN SITU HYBRIDIZATION
| Autor: | Jan M Ruijter, Jaco Hagoort, Piet AJ De Boer, Antoon FM Moorman |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: | |
| Zdroj: | Image Analysis and Stereology, Vol 20, Iss 3, Pp 219-224 (2011) |
| Druh dokumentu: | article |
| ISSN: | 1580-3139 1854-5165 |
| DOI: | 10.5566/ias.v20.p219-224 |
| Popis: | Densitometry on autoradiographs of sections processed for in situ hybridization provides a direct measure for the in situ quantification of mRNA. Gelatin spots, containing different concentrations of the radioisotope, and processed in parallel with the tissue sections, can be used as a sensitive model to calibrate the densitometric measurements. The shape of the gelatin spots was shown to be circular with a parabolic crosssectional profile. This simple shape allows the subdivision of the spot into a series of concentric rings, which enables an unbiased measurement of the optical density - radioactivity relation. This spot measurement is also applicable to DNA arrays spotted on glass or membranes. A new model, explaining the optical density of autoradiographs, was derived and fitted to the calibration points. The use of this calibration method is crucial for the correct interpretation of autoradiographs |
| Databáze: | Directory of Open Access Journals |
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