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Localization of HPV and SC35 Domains in SiHa and CaSki Cells by TSA Using Fluorescent Tyramides

Published online by Cambridge University Press:  02 July 2020

K.E. Adler ,
P.T. Moen Jr. ,
T.J. Erickson  and
M.N. Bobrow
K.E. Adler
Affiliation:
NEN Life Science Products, Boston, MA
P.T. Moen Jr.
Affiliation:
NEN Life Science Products, Boston, MA
T.J. Erickson
Affiliation:
NEN Life Science Products, Boston, MA
M.N. Bobrow
Affiliation:
NEN Life Science Products, Boston, MA
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Extract

Detection of integrated HPV DNA in SiHa and CaSki cells has been demonstrated by in situ PCR (1,2,3) and more recently TSA (4). These reports focused on high sensitivity single copy HPV detection using chromogenic techniques. Here we utilize high resolution and high sensitivity fluorescent methods of detection to analyze the spatial relationship of HPV DNA with respect to subnuclear domains enriched in splicing-related proteins, shown here by immunofluorescent detection of the SC-35 antigen (5). In situhybridization was performed using conventional reagents and the resulting probe signal amplified using fluorescent labeled tyramides (6). In contrast to techniques such as in situ PCR which require specialized procedures and equipment, detection of HPV DNA in SiHa cells was rapidly and routinely achieved using no specialized equipment. In addition, high target resolution was maintained, in contrast to in situ PCR, which is plagued by signal dispersion and difficult localization (7).

Type
Cytochemistry, Histochemistry, Immunocytochemistry, and in Situ Hybridization
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 141 - 142
Copyright
Copyright © Microscopy Society of America 1997

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References

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