In situ hybridization

Abstract

In situ hybridization (ISH) is employed principally to detect mRNA within cells and tissues. ISH begins with the synthesis of a nucleic acid probe complementary in sequence to a cellular target. Through incorporation of a radioactive nucleotide into the probe, a cellular target may be visualized using autoradiography. A nonradioactive variation of this technique utilizing digoxigenin has been championed by some researchers, but our experience has been that radioactive probes produce more reliable and accurate results. Although not a difficult technique, ISH incorporates many steps, which must be performed with care and precision to obtain useful results.