This tutorial paper will review radioactive and non-radioactive in situ hybridization (ISH) techniques at the electron microscope (EM) level; describe our efforts in comparison to those of other groups who recently have published in this field and discuss some potential future applications. Our contribution is the development of a non-radioactive, postembedding technique for the detection of transcripts on thin sections of Lowicryl K4M-embedded cells or tissues. Biotinylated probes were prepared by nick-translation. Signal detection was accomplished with anti-biotin antibody and protein A-gold complexes. Specific labeling was obtained over structures known to be the site of expression. Compared to EM autoradiography this technique offers advantages: (a) rapid signal detection; (b) superior morphological preservation and (c) superior spatial resolution.
"In Situ Hybridization at the Electron Microscope Level,"
Scanning Microscopy: Vol. 1
, Article 30.
Available at: https://digitalcommons.usu.edu/microscopy/vol1/iss1/30