Determination of optimum formalin fixation duration for prostate needle biopsies for immunohistochemistry and quantum dot FISH analysis

Ubaradka G. Sathyanarayana, Chandler Birch, Raymond B Nagle, Scott A. Tomlins, Nallasivam Palanisamy, Wenjun Zhang, Antony Hubbard, Patrick Brunhoeber, Yixin Wang, Lei Tang

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Prostate biopsy is the key clinical specimen for disease diagnosis. However, various conditions used during biopsy processing for histologic analysis may affect the performance of diagnostic tests, such as hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), or in situ hybridization (ISH). One such condition that may affect diagnostic test performance is fixation duration in 10% neutral buffered formalin (NBF). For example, prostate needle biopsies are often <1 mm in diameter and thus overfixed. It is important to understand the impact of tissue fixation duration on diagnostic test performance to enable optimized assay procedures. This study was designed to study the effect of 10% NBF fixation duration of prostate needle biopsy on multiplexed quantum dot (QD) ISH assay of ERG and PTEN, 2 genes commonly altered in prostate cancer. The samples were also evaluated for H&E staining and ERG and PTEN IHC. H&E staining and ERG and PTEN IHC were acceptable for all the durations of fixation tested. For QD ISH, we observed good signals with biopsy samples fixed from 4 to 120 hours. Biopsy specimens fixed between 8 and 72 hours gave the best signal as scored by the study pathologist. In a separate cohort of 18 routinely processed prostate biopsy cores, all cores were stained successfully with the QD ISH assay, and results were 100% concordant to ERG and PTEN IHC. We conclude that 8 to 72 hours duration of fixation for prostate needle biopsies in 10% NBF results in optimal QD ISH assay performance.

Original languageEnglish (US)
Pages (from-to)364-373
Number of pages10
JournalApplied Immunohistochemistry and Molecular Morphology
Volume23
Issue number5
DOIs
StatePublished - May 26 2015

Fingerprint

Quantum Dots
Needle Biopsy
Formaldehyde
In Situ Hybridization
Prostate
Immunohistochemistry
Biopsy
Routine Diagnostic Tests
Staining and Labeling
Tissue Fixation
Hematoxylin
Eosine Yellowish-(YS)
Prostatic Neoplasms
Genes

Keywords

  • formalin fixed paraffin embedded
  • gene expression
  • IHC
  • prostate cancer
  • quantum dot
  • tissue diagnostic

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Medical Laboratory Technology
  • Histology

Cite this

Determination of optimum formalin fixation duration for prostate needle biopsies for immunohistochemistry and quantum dot FISH analysis. / Sathyanarayana, Ubaradka G.; Birch, Chandler; Nagle, Raymond B; Tomlins, Scott A.; Palanisamy, Nallasivam; Zhang, Wenjun; Hubbard, Antony; Brunhoeber, Patrick; Wang, Yixin; Tang, Lei.

In: Applied Immunohistochemistry and Molecular Morphology, Vol. 23, No. 5, 26.05.2015, p. 364-373.

Research output: Contribution to journalArticle

Sathyanarayana, Ubaradka G. ; Birch, Chandler ; Nagle, Raymond B ; Tomlins, Scott A. ; Palanisamy, Nallasivam ; Zhang, Wenjun ; Hubbard, Antony ; Brunhoeber, Patrick ; Wang, Yixin ; Tang, Lei. / Determination of optimum formalin fixation duration for prostate needle biopsies for immunohistochemistry and quantum dot FISH analysis. In: Applied Immunohistochemistry and Molecular Morphology. 2015 ; Vol. 23, No. 5. pp. 364-373.
@article{2e6abc9657dc4e22b4ac24164c8d687e,
title = "Determination of optimum formalin fixation duration for prostate needle biopsies for immunohistochemistry and quantum dot FISH analysis",
abstract = "Prostate biopsy is the key clinical specimen for disease diagnosis. However, various conditions used during biopsy processing for histologic analysis may affect the performance of diagnostic tests, such as hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), or in situ hybridization (ISH). One such condition that may affect diagnostic test performance is fixation duration in 10{\%} neutral buffered formalin (NBF). For example, prostate needle biopsies are often <1 mm in diameter and thus overfixed. It is important to understand the impact of tissue fixation duration on diagnostic test performance to enable optimized assay procedures. This study was designed to study the effect of 10{\%} NBF fixation duration of prostate needle biopsy on multiplexed quantum dot (QD) ISH assay of ERG and PTEN, 2 genes commonly altered in prostate cancer. The samples were also evaluated for H&E staining and ERG and PTEN IHC. H&E staining and ERG and PTEN IHC were acceptable for all the durations of fixation tested. For QD ISH, we observed good signals with biopsy samples fixed from 4 to 120 hours. Biopsy specimens fixed between 8 and 72 hours gave the best signal as scored by the study pathologist. In a separate cohort of 18 routinely processed prostate biopsy cores, all cores were stained successfully with the QD ISH assay, and results were 100{\%} concordant to ERG and PTEN IHC. We conclude that 8 to 72 hours duration of fixation for prostate needle biopsies in 10{\%} NBF results in optimal QD ISH assay performance.",
keywords = "formalin fixed paraffin embedded, gene expression, IHC, prostate cancer, quantum dot, tissue diagnostic",
author = "Sathyanarayana, {Ubaradka G.} and Chandler Birch and Nagle, {Raymond B} and Tomlins, {Scott A.} and Nallasivam Palanisamy and Wenjun Zhang and Antony Hubbard and Patrick Brunhoeber and Yixin Wang and Lei Tang",
year = "2015",
month = "5",
day = "26",
doi = "10.1097/PAI.0000000000000085",
language = "English (US)",
volume = "23",
pages = "364--373",
journal = "Applied Immunohistochemistry and Molecular Morphology",
issn = "1541-2016",
publisher = "Lippincott Williams and Wilkins",
number = "5",

}

TY - JOUR

T1 - Determination of optimum formalin fixation duration for prostate needle biopsies for immunohistochemistry and quantum dot FISH analysis

AU - Sathyanarayana, Ubaradka G.

AU - Birch, Chandler

AU - Nagle, Raymond B

AU - Tomlins, Scott A.

AU - Palanisamy, Nallasivam

AU - Zhang, Wenjun

AU - Hubbard, Antony

AU - Brunhoeber, Patrick

AU - Wang, Yixin

AU - Tang, Lei

PY - 2015/5/26

Y1 - 2015/5/26

N2 - Prostate biopsy is the key clinical specimen for disease diagnosis. However, various conditions used during biopsy processing for histologic analysis may affect the performance of diagnostic tests, such as hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), or in situ hybridization (ISH). One such condition that may affect diagnostic test performance is fixation duration in 10% neutral buffered formalin (NBF). For example, prostate needle biopsies are often <1 mm in diameter and thus overfixed. It is important to understand the impact of tissue fixation duration on diagnostic test performance to enable optimized assay procedures. This study was designed to study the effect of 10% NBF fixation duration of prostate needle biopsy on multiplexed quantum dot (QD) ISH assay of ERG and PTEN, 2 genes commonly altered in prostate cancer. The samples were also evaluated for H&E staining and ERG and PTEN IHC. H&E staining and ERG and PTEN IHC were acceptable for all the durations of fixation tested. For QD ISH, we observed good signals with biopsy samples fixed from 4 to 120 hours. Biopsy specimens fixed between 8 and 72 hours gave the best signal as scored by the study pathologist. In a separate cohort of 18 routinely processed prostate biopsy cores, all cores were stained successfully with the QD ISH assay, and results were 100% concordant to ERG and PTEN IHC. We conclude that 8 to 72 hours duration of fixation for prostate needle biopsies in 10% NBF results in optimal QD ISH assay performance.

AB - Prostate biopsy is the key clinical specimen for disease diagnosis. However, various conditions used during biopsy processing for histologic analysis may affect the performance of diagnostic tests, such as hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), or in situ hybridization (ISH). One such condition that may affect diagnostic test performance is fixation duration in 10% neutral buffered formalin (NBF). For example, prostate needle biopsies are often <1 mm in diameter and thus overfixed. It is important to understand the impact of tissue fixation duration on diagnostic test performance to enable optimized assay procedures. This study was designed to study the effect of 10% NBF fixation duration of prostate needle biopsy on multiplexed quantum dot (QD) ISH assay of ERG and PTEN, 2 genes commonly altered in prostate cancer. The samples were also evaluated for H&E staining and ERG and PTEN IHC. H&E staining and ERG and PTEN IHC were acceptable for all the durations of fixation tested. For QD ISH, we observed good signals with biopsy samples fixed from 4 to 120 hours. Biopsy specimens fixed between 8 and 72 hours gave the best signal as scored by the study pathologist. In a separate cohort of 18 routinely processed prostate biopsy cores, all cores were stained successfully with the QD ISH assay, and results were 100% concordant to ERG and PTEN IHC. We conclude that 8 to 72 hours duration of fixation for prostate needle biopsies in 10% NBF results in optimal QD ISH assay performance.

KW - formalin fixed paraffin embedded

KW - gene expression

KW - IHC

KW - prostate cancer

KW - quantum dot

KW - tissue diagnostic

UR - http://www.scopus.com/inward/record.url?scp=84929841121&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84929841121&partnerID=8YFLogxK

U2 - 10.1097/PAI.0000000000000085

DO - 10.1097/PAI.0000000000000085

M3 - Article

VL - 23

SP - 364

EP - 373

JO - Applied Immunohistochemistry and Molecular Morphology

JF - Applied Immunohistochemistry and Molecular Morphology

SN - 1541-2016

IS - 5

ER -