Document Detail


Application of confocal laser scanning microscopy (CLSM) to visualize prolactin (PRL) and PRL mRNA in the normal and estrogen-treated rat pituitary glands using non-fluorescent probes.
MedLine Citation:
PMID:  9361267     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In the present study, we performed concomitant visualization of immunohistochemistry (IHC) and in situ hybridization (ISH) on the materials processed for conventional light microscopic specimens using non-fluorescent Confocal Laser. Scanning Microscopy (CLSM). CLSM was used in the reflection confocal mode using horseradish peroxidase (HRP)-3-3'diaminobenzidine (DAB)-osmium (osmium black) and nitroblue tetrazolium (NBT) as non-fluorescent detection methods (probes). To obtain clearer images of the organelles, images that were built up as electronic signals in CLSM were processed in an image analysis system (IAS). By using the combination of CLSM and IAS, in IHC, immunohistochemical localization of prolactin (PRL) was in well-developed lamellar or whorling rough endoplasmic reticula (RER), Golgi apparatus, and secretory granules. With ISH, the expression and distribution of PRL messenger ribonucleic acid (mRNA) was observed in a fashion suggesting polysome-like structures on RER. These observations were confirmed by immunoelectron microscopy and electron microscopic ISH. The herein-described method is expected to be useful to perform the concomitant observation of IHC and ISH at subcellular levels using the conventional light microscopic specimens.
Authors:
J Itoh; N Sanno; A Matsuno; Y Itoh; K Watanabe; R Y Osamura
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Microscopy research and technique     Volume:  39     ISSN:  1059-910X     ISO Abbreviation:  Microsc. Res. Tech.     Publication Date:  1997 Oct 
Date Detail:
Created Date:  1997-12-17     Completed Date:  1997-12-17     Revised Date:  2003-11-14    
Medline Journal Info:
Nlm Unique ID:  9203012     Medline TA:  Microsc Res Tech     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  157-67     Citation Subset:  IM    
Affiliation:
Laboratories for Structure and Function Research, School of Medicine, Tokai University, Kanagawa, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Cytoplasmic Granules / metabolism
Endoplasmic Reticulum, Rough / metabolism
Estrogens / pharmacology*
Female
Golgi Apparatus / metabolism
Image Processing, Computer-Assisted
Immunohistochemistry / methods
In Situ Hybridization
Lasers*
Microscopy, Confocal / methods*
Microscopy, Electron
Microscopy, Immunoelectron
Pituitary Gland / cytology,  metabolism*,  ultrastructure*
Prolactin / analysis*
RNA, Messenger / analysis*
Rats
Rats, Wistar
Chemical
Reg. No./Substance:
0/Estrogens; 0/RNA, Messenger; 9002-62-4/Prolactin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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