| Freeze substitution and freeze drying for stable, long-term preservation of cytologic specimens for immunostaining. | |
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MedLine Citation:
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PMID: 8669168 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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OBJECTIVE: To develop a new method of fixing and preserving cytologic specimens for immunostaining after long-term storage at room temperature. STUDY DESIGN: The method consists of three steps: fixation, freeze substitution or freeze drying, and storage. To test the method, we used the human small cell lung carcinoma cell line Lu-135, which expresses a high level of mutant p53 protein and exhibits strong nuclear staining when reacted with an anti-p53 antibody. A smear of Lu-135 cells was fixed in a mixture of methanol and ether (50:50=vol/vol) or sprayed with a fixative containing isopropyl alcohol, methanol and polyethylene glycol. The fixed cells were freeze substituted by immersing them in a dry ice/methanol/ether bath and then were freeze dried under a vacuum. The smear was then placed in a 50-mL conical tube containing silica gel. The tube was sealed and stored at room temperature. RESULTS: Freeze substituted cells that were fixed with methanol/ether and stored for more than six months retained strong p53 positivity, as strong as that of the control cells, which had been fixed and stored in methanol. CONCLUSION: Freeze substitution and freeze drying are an alternative method of preserving cytologic specimens. |
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Authors:
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R Takahashi; T Okuyama; S Matsuo; S Maeda |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Acta cytologica Volume: 40 ISSN: 0001-5547 ISO Abbreviation: Acta Cytol. Publication Date: 1996 May-Jun |
Date Detail:
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Created Date: 1996-08-05 Completed Date: 1996-08-05 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0370307 Medline TA: Acta Cytol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 396-400 Citation Subset: IM |
Affiliation:
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Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Japan. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Carcinoma, Small Cell
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diagnosis Cytological Techniques* Desiccation Freeze Drying / methods* Freeze Substitution / methods* Humans Immunohistochemistry Nitrogen Oxygen Specimen Handling / methods* Staining and Labeling / methods Tissue Fixation / methods Tumor Cells, Cultured / cytology |
| Chemical | |
Reg. No./Substance:
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7727-37-9/Nitrogen; 7782-44-7/Oxygen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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