Fundamentals of quantitative autoradiography by computer densitometry for in situ hybridization, with emphasis on 33P

The goal of quantitative autoradiography (QAR) in in situ hybridization (ISH) is to determine the amount of radioactive oligonucleotide or riboprobe present in the corresponding area of the tissue slice that produced an autoradiographic image. This article discusses (a) some of the considerations related to selection and use of computer image analysis systems for accomplishing this objective, (b) development of 14C plastic autoradiographic standards for ISH QAR with 33P, (c) using QAR to develop Tm curves for ISH probes, and (d) measurement of resolution with video imaging systems for QAR.