Epitope Preservation Methods for Tissue Microarrays

Nicole Andeen, Regina Bowman, Toni Baullinger, J. Mathew Brooks, Maria S. Tretiakova

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)380-389
Number of pages10
JournalAmerican journal of clinical pathology
Volume148
Issue number5
DOIs
StatePublished - Nov 1 2017
Externally publishedYes

Fingerprint

Tissue Preservation
Vacuum
Paraffin
Epitopes
Antibodies
Isocitrate Dehydrogenase
Synaptophysin
Temperature
Androgen Receptors
Air
Guidelines

Keywords

  • Antibody
  • Antigenicity
  • Epitope
  • Immunoreactivity
  • Storage
  • Tissue microarray

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Andeen, N., Bowman, R., Baullinger, T., Brooks, J. M., & Tretiakova, M. S. (2017). Epitope Preservation Methods for Tissue Microarrays. American journal of clinical pathology, 148(5), 380-389. https://doi.org/10.1093/ajcp/aqx062

Epitope Preservation Methods for Tissue Microarrays. / Andeen, Nicole; Bowman, Regina; Baullinger, Toni; Brooks, J. Mathew; Tretiakova, Maria S.

In: American journal of clinical pathology, Vol. 148, No. 5, 01.11.2017, p. 380-389.

Research output: Contribution to journalArticle

Andeen, N, Bowman, R, Baullinger, T, Brooks, JM & Tretiakova, MS 2017, 'Epitope Preservation Methods for Tissue Microarrays', American journal of clinical pathology, vol. 148, no. 5, pp. 380-389. https://doi.org/10.1093/ajcp/aqx062
Andeen, Nicole ; Bowman, Regina ; Baullinger, Toni ; Brooks, J. Mathew ; Tretiakova, Maria S. / Epitope Preservation Methods for Tissue Microarrays. In: American journal of clinical pathology. 2017 ; Vol. 148, No. 5. pp. 380-389.
@article{77c0d4dfe7ae4c2c934f5121d90e0727,
title = "Epitope Preservation Methods for Tissue Microarrays",
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.",
keywords = "Antibody, Antigenicity, Epitope, Immunoreactivity, Storage, Tissue microarray",
author = "Nicole Andeen and Regina Bowman and Toni Baullinger and Brooks, {J. Mathew} and Tretiakova, {Maria S.}",
year = "2017",
month = "11",
day = "1",
doi = "10.1093/ajcp/aqx062",
language = "English (US)",
volume = "148",
pages = "380--389",
journal = "American Journal of Clinical Pathology",
issn = "0002-9173",
publisher = "American Society of Clinical Pathologists",
number = "5",

}

TY - JOUR

T1 - Epitope Preservation Methods for Tissue Microarrays

AU - Andeen, Nicole

AU - Bowman, Regina

AU - Baullinger, Toni

AU - Brooks, J. Mathew

AU - Tretiakova, Maria S.

PY - 2017/11/1

Y1 - 2017/11/1

N2 - 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.

AB - 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.

KW - Antibody

KW - Antigenicity

KW - Epitope

KW - Immunoreactivity

KW - Storage

KW - Tissue microarray

UR - http://www.scopus.com/inward/record.url?scp=85033554854&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85033554854&partnerID=8YFLogxK

U2 - 10.1093/ajcp/aqx062

DO - 10.1093/ajcp/aqx062

M3 - Article

VL - 148

SP - 380

EP - 389

JO - American Journal of Clinical Pathology

JF - American Journal of Clinical Pathology

SN - 0002-9173

IS - 5

ER -