Document Detail


Addition of phosphotungstic acid to ethanol for dehydration improves both the ultrastructure and antigenicity of pituitary tissue embedded in LR White acrylic resin.
MedLine Citation:
PMID:  16505580     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Although hydrophilic acrylic resins including LR White have been widely utilized as embedding media for immunocytochemical use, the constituents of tissues are often extracted by the resin monomer during the infiltration process of the embedment, resulting in a discernible impairment of the ultrastructure when the tissue is weakly fixed only with aldehydes. To minimize the extraction by the resin monomer, the embedding procedure with LR White resin was reexamined in the present study. Among the treatments tested, a partial dehydration with 70% ethanol containing 2% phosphotungstic acid (PTA) well preserved the ultrastructure of the pituitary tissue without spoiling the antigenicity of LHbeta and other representative markers for the Golgi apparatus. In addition, treatment with 1% tannic acid (TA) prior to the dehydration described above synergistically improved both the ultrastructure and antigenicity of the tissue so that the orientation of the Golgi apparatus could be determined by double immunogold labeling with commercially available anti-GM130 and anti-TGN38 antibodies. The ultrathin sections from the LR White-embedded tissue treated with TA and dehydrated in 70% ethanol containing 2% PTA also enhanced contrast without conventional heavy-metal staining with uranyl acetate and lead citrate. Our findings further suggest that the precipitation of TA and PTA protected the tissue from being extracted during the embedment, probably because an insoluble complex was transiently formed with the constituents of the tissue. This simple modification of the LR White embedment can extend the application of post-embedding immunocytochemistry as an alternative to pre-embedding immunolabeling with frozen ultrathin sections.
Authors:
Yuko Sakai; Masahiro Hosaka; Yoshiki Hira; Tsuyoshi Watanabe
Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Archives of histology and cytology     Volume:  68     ISSN:  0914-9465     ISO Abbreviation:  Arch. Histol. Cytol.     Publication Date:  2005 Dec 
Date Detail:
Created Date:  2006-02-28     Completed Date:  2006-03-27     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8806082     Medline TA:  Arch Histol Cytol     Country:  Japan    
Other Details:
Languages:  eng     Pagination:  337-47     Citation Subset:  IM    
Affiliation:
Department of Anatomy II, Asahikawa Medical College, Midorigaoka-higashi 2-1-1-1, Asahikawa 078-8510, Japan.
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MeSH Terms
Descriptor/Qualifier:
Acrylic Resins*
Animals
Antigens / immunology*
Desiccation
Ethanol / chemistry*
Immunohistochemistry
Luteinizing Hormone, beta Subunit / metabolism
Male
Phosphotungstic Acid / chemistry*
Pituitary Gland / immunology*,  ultrastructure*
Rats
Rats, Wistar
Tissue Embedding
Tissue Fixation
Chemical
Reg. No./Substance:
0/Acrylic Resins; 0/Antigens; 0/Luteinizing Hormone, beta Subunit; 12067-99-1/Phosphotungstic Acid; 64-17-5/Ethanol; 94188-59-7/LR white

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


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