Document Detail

Freeze substitution and freeze drying for stable, long-term preservation of cytologic specimens for immunostaining.
MedLine Citation:
PMID:  8669168     Owner:  NLM     Status:  MEDLINE    
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.
R Takahashi; T Okuyama; S Matsuo; S Maeda
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Acta cytologica     Volume:  40     ISSN:  0001-5547     ISO Abbreviation:  Acta Cytol.     Publication Date:    1996 May-Jun
Date Detail:
Created Date:  1996-08-05     Completed Date:  1996-08-05     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0370307     Medline TA:  Acta Cytol     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  396-400     Citation Subset:  IM    
Department of Pathology and Tumor Biology, Graduate School of Medicine, Kyoto University, Japan.
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MeSH Terms
Carcinoma, Small Cell / diagnosis
Cytological Techniques*
Freeze Drying / methods*
Freeze Substitution / methods*
Specimen Handling / methods*
Staining and Labeling / methods
Tissue Fixation / methods
Tumor Cells, Cultured / cytology
Reg. No./Substance:
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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