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.
authors.affiliation: *Ventana Medical Systems †Tissue Acquisition and Cellular/Molecular Analysis Shared Service (TACMASS), University of Arizona Cancer Center, Tucson, AZ ‡Department of Pathology, Michigan Center for Translational Pathology §Department of Urology ∥Comprehensive Cancer Center, University of Michigan Medical School, Ann Arbor, MI.
authors.forename: Ubaradka G
authors.initials: UG
authors.lastname: Sathyanarayana
authors.forename: Chandler
authors.initials: C
authors.lastname: Birch
authors.forename: Raymond B
authors.initials: RB
authors.lastname: Nagle
authors.forename: Scott A
authors.initials: SA
authors.lastname: Tomlins
authors.forename: Nallasivam
authors.initials: N
authors.lastname: Palanisamy
authors.forename: Wenjun
authors.initials: W
authors.lastname: Zhang
authors.forename: Antony
authors.initials: A
authors.lastname: Hubbard
authors.forename: Patrick
authors.initials: P
authors.lastname: Brunhoeber
authors.forename: Yixin
authors.initials: Y
authors.lastname: Wang
authors.forename: Lei
authors.initials: L
authors.lastname: Tang
mesh_terms: D000230:Adenocarcinoma / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology / Q000601:surgery; D014408:Biomarkers, Tumor / Q000235:genetics*; D001707:Biopsy, Needle; D015331:Cohort Studies; D004801:Eosine Yellowish-(YS); D005557:Formaldehyde; D015870:Gene Expression; D006416:Hematoxylin; D006801:Humans; D007150:Immunohistochemistry / Q000592:standards*; D017404:In Situ Hybridization, Fluorescence / Q000592:standards*; D008297:Male; D051059:PTEN Phosphohydrolase / Q000235:genetics; D011467:Prostate / Q000378:metabolism / Q000473:pathology / Q000601:surgery; D011471:Prostatic Neoplasms / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology / Q000601:surgery; D045663:Quantum Dots; D016610:Tissue Embedding; D016707:Tissue Fixation / Q000379:methods*; D015534:Trans-Activators / Q000235:genetics; D000071230:Transcriptional Regulator ERG
chemical_list: D014408:Biomarkers, Tumor; C072312:ERG protein, human; D015534:Trans-Activators; D000071230:Transcriptional Regulator ERG; D005557:Formaldehyde; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human; D004801:Eosine Yellowish-(YS); D006416:Hematoxylin