In situ hybridization at the electron microscope level.
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: rapid signal detection; superior morphological preservation and superior spatial resolution.
D000818:Animals; D004275:DNA, Ribosomal / Q000235:genetics; D004330:Drosophila; D005260:Female; D008854:Microscopy, Electron / Q000379:methods; D008928:Mitochondria / Q000648:ultrastructure; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization; D010053:Ovary / Q000166:cytology / Q000648:ultrastructure; D014176:Protein Biosynthesis; D014316:Tritium
D004275:DNA, Ribosomal; D014316:Tritium