Hostname: page-component-cd9895bd7-mkpzs Total loading time: 0 Render date: 2024-12-25T19:07:38.755Z Has data issue: false hasContentIssue false
  • English
  • Français

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
Get access

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

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

1) Nuovo, G.J.et al., Am J Pathol, 139(1991)847Google Scholar

2) Zehbe, I.et al., Anticancer Res, 12(1992)2165Google Scholar

3) O’Leary, J.J.et al., J Clin pathol 47(1994)933CrossRefGoogle Scholar

4) Zehbe, I., et al., (in press), J Cell VisionGoogle Scholar

5) XD, Fuet al., Proc Natl Acad Sci USA 89(1992)1725Google Scholar

6) Raap, A.K.et al., Human Mol Genet, 4 #4 (1995)529CrossRefGoogle Scholar

7) Haase, A.T.et al., Proc Natl Acad Sci USA, 87(1990)4971CrossRefGoogle Scholar

8) Moen, P.T. Jr, et al., Hum Mol Genet 4 Spec (1995)1779CrossRefGoogle Scholar

9) Sandi-Goldin, R.M.et al., J Virol, 69(1995)6063CrossRefGoogle Scholar