Nicole K Andeen 1 , Regina Bowman 1 , Toni Baullinger 1 , J Mathew Brooks 1 , Maria S Tretiakova 1 . Show Affiliations »
Abstract
OBJECTIVES: We sought to test recent guidelines for preserving immunoreactivity of precut slides, to quantify loss of immunoreactivity, and to determine potential for preservation by altering storage conditions. METHODS: Precut slides from tissue microarrays were stored under one of several conditions: exposed to ambient air at room temperature, 4°C, or -20°C or in a vacuum-sealed container at room temperature, -20°C, -80°C, or with paraffin coating. At multiple intervals over 1 year, slides were stained with antibodies against p53, isocitrate dehydrogenase 1, Ki-67, synaptophysin, and androgen receptor and evaluated. RESULTS: Compared with time 0, the overall median percentage immunoreactivity was 66% at 6 months and 51% at 1 year. During the experiment, this was as low as 55% for precut slides stored in paraffin coating and up to 87% for those stored at -20°C. Vacuum sealing was an effective preservative for some antibody targets and detrimental for others. Storage at -80°C did not have added value. CONCLUSIONS: For precut slides, there is a time, storage condition, and antibody-dependent loss of immunoreactivity that could compromise analysis of prognostic, predictive, and diagnostic markers. Our findings support previous recommendations and suggest that the best storage conditions are at -20°C, without paraffin coating or vacuum sealing. © American Society for Clinical Pathology, 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com
OBJECTIVES: We sought to test recent guidelines for preserving immunoreactivity of precut slides, to quantify loss of immunoreactivity, and to determine potential for preservation by altering storage conditions. METHODS: Precut slides from tissue microarrays were stored under one of several conditions: exposed to ambient air at room temperature, 4°C, or -20°C or in a vacuum-sealed container at room temperature, -20°C, -80°C, or with paraffin coating. At multiple intervals over 1 year, slides were stained with antibodies against p53, isocitrate dehydrogenase 1, Ki-67, synaptophysin, and androgen receptor and evaluated. RESULTS: Compared with time 0, the overall median percentage immunoreactivity was 66% at 6 months and 51% at 1 year. During the experiment, this was as low as 55% for precut slides stored in paraffin coating and up to 87% for those stored at -20°C. Vacuum sealing was an effective preservative for some antibody targets and detrimental for others. Storage at -80°C did not have added value. CONCLUSIONS: For precut slides, there is a time, storage condition, and antibody-dependent loss of immunoreactivity that could compromise analysis of prognostic, predictive, and diagnostic markers. Our findings support previous recommendations and suggest that the best storage conditions are at -20°C, without paraffin coating or vacuum sealing. © American Society for Clinical Pathology, 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com
Entities: Chemical
Keywords:
Antibody; Antigenicity; Epitope; Immunoreactivity; Storage; Tissue microarray
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Substances: See more »
Year: 2017
PMID: 29106459 PMCID: PMC6951944 DOI: 10.1093/ajcp/aqx062
Source DB: PubMed Journal: Am J Clin Pathol ISSN: 0002-9173 Impact factor: 2.493