DNA Labeling at Electron Microscopy.
Here, we describe a method for locating DNA on ultrathin sections. This technique is compatible with all usual fixation and embedding procedures and can be combined with cytochemical methods. Ultrathin sections are incubated in a medium containing terminal deoxynucleotidyl transferase (TdT) and various non-isotopic nucleotide analogs. The labeled nucleotides bound to the surface of ultrathin sections are then visualized by an indirect immunogold labeling technique. This high-resolution method provides a powerful tool for pinpointing the precise location of DNA within biological material, even where DNA is present in very low amounts.
Giga Neurosciences, Department of Life Sciences, Laboratory of Cell Biology, University of Liège, Quartier Hôpital, 15 Avenue Hippocrate (B36), 4000, Liege, Belgium.
Nicolas
N
Thelen
Giga Neurosciences, Department of Life Sciences, Laboratory of Cell Biology, University of Liège, Quartier Hôpital, 15 Avenue Hippocrate (B36), 4000, Liege, Belgium. mthiry@ulg.ac.be.
Marc
M
Thiry
D004247:DNA / Q000235:genetics; D007150:Immunohistochemistry; D008854:Microscopy, Electron / Q000379:methods*; D016253:Microscopy, Immunoelectron / Q000379:methods; D013194:Staining and Labeling / Q000379:methods
D004247:DNA
DNA labeling; Immuno-electron microscopy; Immunocytochemistry; Immunogold techniques; Terminal deoxynucleotidyl transferase