Investigation of tissue preparation conditions for non-radioactive in situ hybridization: localization of transforming growth factor-alpha message in rat kidney.
A non-radioactive method of in situ hybridization was used to localize transforming growth factor-alpha mRNA in epithelial cells of collecting ducts and tubules in rat kidney tissue sections. The intensity and specificity of staining were assessed under a variety of tissue preparation conditions, including a direct comparison of paraffin against frozen sections. Under optimal conditions, both the signal strength and the cellular localization of the growth factor message were superior in paraffin sections. The staining method could also be used to localize the message in lung tissue, indicating that the procedure is generally applicable to other tissues. Our results indicate that the use of paraffin sections for nonradioactive in situ hybridization affords a number of advantages for the localization of specific messages in tissue sections.
Liu
J Y
JY
Department of Pathology, Tulane University Medical Center, New Orleans, LA 70112, USA.
Lei
W H
WH
Brody
A R
AR
Hoyle
G W
GW
D000818:Animals; D015925:Cryopreservation; D004847:Epithelial Cells / Q000378:metabolism; D017403:In Situ Hybridization; D007668:Kidney / Q000166:cytology / Q000378:metabolism*; D007684:Kidney Tubules / Q000166:cytology / Q000378:metabolism; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000378:metabolism; D008168:Lung / Q000378:metabolism; D016612:Paraffin Embedding; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D016211:Transforming Growth Factor alpha / Q000235:genetics / Q000378:metabolism*