Localization of HPV-16 DNA sequence in CaSki cells by electron microscopic hybridocytochemistry.

Abstract

We investigated the HPV-16 DNA sequence in the CaSki cervical carcinoma cell line by electron microscopic hybridocytochemistry using biotinylated HPV-16/18 probes. At the light microscopic level, reaction product of hybridized HPV-16 DNA sequence was not seen in the cytoplasm but appeared as spots or rods randomly distributed in the nuclei. By electron miaoscopy, reaction product was seen aggregated in several regions in the nuclei. Most of the stained areas did not reveal particular architecture but showed part of the chromatin structure. In other nuclei, reaction product was observed to be associated with strings of loop-like structure, and some stained loops were seen to be connected directly

Biotinylated probes for in situ hybridization histochemistry: use for mRNA detection.

“…The use of streptavidin linked to alkaline phosphatase with bromochloro-indolyl phosphate and nitroblue tetrazolium as the substrate is presently one ofthe most efficient and sensitive ways to detect "As with biotinylated probes at the LM level ( 6.9,10,12,19,21,25,26). Gold particles or peroxidase provide exquisite subcellular resolution for EM (2, 16,18,27).…”

“…This is especially important for detecting mRNAs in neurons ( Figure 1B) and other highly compartmentalized cells, where localization of radioactive probes is less precise (4). Biotinylated probes can be used with cryostat or Vibratome sections (4, 9,17), cultured cells (14,23,24), and tissues embedded in paraffin or plastic (2,5,6,10, 16,18,27). It appears that a biotinylated probe hybridized to its corresponding mRNA is unaffected by plastic-embedding procedures and can be detected in semi-thin plastic sections, thus providing optimal subcellular resolution of the signal and the possibility of correlative immunohistochemistry (10) (Figures lC, lD, and IF).…”