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. 3
Publication year     2001
Volume     str. 219-24
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 measurements. The shape of the gelatin spots was shown to be circular with a parabolic cross-sectional profile. This sample shape allows the subdivision of the spot into 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     CALIBRATION
DENSITOMETRY
IN SITU HYBRIDIZATION
AUTORADIOGRAPHY
RNA, MESSENGER
GELATIN