Evaluation of Optimized Toluidine Blue Stain as an Alternative Stain for Rapid On-Site Evaluation (ROSE)

<b>Background</b>: Rapid on-site evaluation (ROSE) is crucial for improving the diagnostic yield of fine-needle aspiration (FNA) biopsies. Despite recent advances in ROSE, such as telecytology, the rapid stains used in this process have not seen significant innovation. Diff-Quik (DQ) and...

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Main Authors: Alejandro S. Mendoza, Alaa Afify, Lydia Howell, John Bishop, Aurelia Lauderdale, Stan Seko, Ronelson Hermosilla, Donald York, Kurt B. Schaberg
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Diagnostics
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Online Access:https://www.mdpi.com/2075-4418/15/10/1223
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author Alejandro S. Mendoza
Alaa Afify
Lydia Howell
John Bishop
Aurelia Lauderdale
Stan Seko
Ronelson Hermosilla
Donald York
Kurt B. Schaberg
author_facet Alejandro S. Mendoza
Alaa Afify
Lydia Howell
John Bishop
Aurelia Lauderdale
Stan Seko
Ronelson Hermosilla
Donald York
Kurt B. Schaberg
author_sort Alejandro S. Mendoza
collection DOAJ
description <b>Background</b>: Rapid on-site evaluation (ROSE) is crucial for improving the diagnostic yield of fine-needle aspiration (FNA) biopsies. Despite recent advances in ROSE, such as telecytology, the rapid stains used in this process have not seen significant innovation. Diff-Quik (DQ) and Toluidine Blue (TB), the most common ROSE stains, have significant limitations. This study evaluates the optimized Toluidine Blue stain, a mixture of <b>T</b>oluidine Blue, <b>E</b>osin, and <b>A</b>lcohol (TEA), as a potential alternative to TB or DQ for ROSE. <b>Methods</b>: A comparative study was conducted using fifty remnant body fluid specimens with adequate cellularity, collected at the University of California Davis Medical Center over six months. Two smears were prepared from each specimen. One was stained with TB, and the other with optimized Toluidine Blue (TEA). Digital images of each slide were evaluated by three cytologists and two cytopathologists, blinded to the stain, using five criteria: background staining, cytoplasmic detail, nuclear membrane clarity, chromatin texture, and nucleoli staining. Each criterion was scored on a scale of 1 to 3. <b>Results</b>: Optimized Toluidine Blue (TEA) stain demonstrated superior overall image quality compared to TB. Specifically, optimized Toluidine Blue (TEA) showed significantly less background staining (<i>p</i> < 0.05) and improved nuclear membrane clarity (<i>p</i> < 0.05), chromatin texture (<i>p</i> < 0.05), and nucleoli detail (<i>p</i> < 0.05). There was no significant difference between the two stains in the assessment of cellularity or cytoplasmic detail. <b>Conclusions</b>: The optimized Toluidine Blue (TEA) stain shows promise as a rapid stain for ROSE, offering rapid processing and improved digital image quality. Further evaluation of optimized Toluidine Blue (TEA) stain on FNA specimens is warranted to validate these findings and explore its potential to enhance telecytology in ROSE.
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spelling doaj-art-6662dca971e343f184357752e902a7be2025-08-20T03:14:31ZengMDPI AGDiagnostics2075-44182025-05-011510122310.3390/diagnostics15101223Evaluation of Optimized Toluidine Blue Stain as an Alternative Stain for Rapid On-Site Evaluation (ROSE)Alejandro S. Mendoza0Alaa Afify1Lydia Howell2John Bishop3Aurelia Lauderdale4Stan Seko5Ronelson Hermosilla6Donald York7Kurt B. Schaberg8Department of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USADepartment of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USADepartment of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USADepartment of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USADepartment of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USADepartment of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USADepartment of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USADepartment of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USADepartment of Pathology and Laboratory Medicine, UC Davis Health, Sacramento, CA 95817, USA<b>Background</b>: Rapid on-site evaluation (ROSE) is crucial for improving the diagnostic yield of fine-needle aspiration (FNA) biopsies. Despite recent advances in ROSE, such as telecytology, the rapid stains used in this process have not seen significant innovation. Diff-Quik (DQ) and Toluidine Blue (TB), the most common ROSE stains, have significant limitations. This study evaluates the optimized Toluidine Blue stain, a mixture of <b>T</b>oluidine Blue, <b>E</b>osin, and <b>A</b>lcohol (TEA), as a potential alternative to TB or DQ for ROSE. <b>Methods</b>: A comparative study was conducted using fifty remnant body fluid specimens with adequate cellularity, collected at the University of California Davis Medical Center over six months. Two smears were prepared from each specimen. One was stained with TB, and the other with optimized Toluidine Blue (TEA). Digital images of each slide were evaluated by three cytologists and two cytopathologists, blinded to the stain, using five criteria: background staining, cytoplasmic detail, nuclear membrane clarity, chromatin texture, and nucleoli staining. Each criterion was scored on a scale of 1 to 3. <b>Results</b>: Optimized Toluidine Blue (TEA) stain demonstrated superior overall image quality compared to TB. Specifically, optimized Toluidine Blue (TEA) showed significantly less background staining (<i>p</i> < 0.05) and improved nuclear membrane clarity (<i>p</i> < 0.05), chromatin texture (<i>p</i> < 0.05), and nucleoli detail (<i>p</i> < 0.05). There was no significant difference between the two stains in the assessment of cellularity or cytoplasmic detail. <b>Conclusions</b>: The optimized Toluidine Blue (TEA) stain shows promise as a rapid stain for ROSE, offering rapid processing and improved digital image quality. Further evaluation of optimized Toluidine Blue (TEA) stain on FNA specimens is warranted to validate these findings and explore its potential to enhance telecytology in ROSE.https://www.mdpi.com/2075-4418/15/10/1223telecytologyFNAROSETEA stainToluidine Blue (TB)Diff-Quik (DQ)
spellingShingle Alejandro S. Mendoza
Alaa Afify
Lydia Howell
John Bishop
Aurelia Lauderdale
Stan Seko
Ronelson Hermosilla
Donald York
Kurt B. Schaberg
Evaluation of Optimized Toluidine Blue Stain as an Alternative Stain for Rapid On-Site Evaluation (ROSE)
Diagnostics
telecytology
FNA
ROSE
TEA stain
Toluidine Blue (TB)
Diff-Quik (DQ)
title Evaluation of Optimized Toluidine Blue Stain as an Alternative Stain for Rapid On-Site Evaluation (ROSE)
title_full Evaluation of Optimized Toluidine Blue Stain as an Alternative Stain for Rapid On-Site Evaluation (ROSE)
title_fullStr Evaluation of Optimized Toluidine Blue Stain as an Alternative Stain for Rapid On-Site Evaluation (ROSE)
title_full_unstemmed Evaluation of Optimized Toluidine Blue Stain as an Alternative Stain for Rapid On-Site Evaluation (ROSE)
title_short Evaluation of Optimized Toluidine Blue Stain as an Alternative Stain for Rapid On-Site Evaluation (ROSE)
title_sort evaluation of optimized toluidine blue stain as an alternative stain for rapid on site evaluation rose
topic telecytology
FNA
ROSE
TEA stain
Toluidine Blue (TB)
Diff-Quik (DQ)
url https://www.mdpi.com/2075-4418/15/10/1223
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