Author/Editor		Ruijter, Jan M; Hagoort, Jaco; de Boer, Piet AJ; Moorman, Antoon FM
Title		Calibration of densitometry in radio-isotopic in situ hybridization
Type		članek
Source		Image Anal Stereol
Vol. and No.		Letnik 20, št. Suppl 1
Publication year		2001
Volume		str. 561-6
Language		eng
Abstract		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 cross-sectional 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
Descriptors		IN SITU HYBRIDIZATION CALIBRATION DENSITOMETRY AUTORADIOGRAPHY IMAGE PROCESSING, COMPUTER-ASSISTED RNA, MESSENGER