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The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue

The use of formalin in histopathological sample preparation is intended to preserve protein and cellular organelles1. It is argued that prolonged formalin fixation may lead to tissue shrinkage and hardening2,3. Reports on immunohistochemical studies showed that this does not cause reduction in antig...

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Main Authors: Mohd Radzuan, Hazulin, Wan Omar, Wan Fatein Nabeila, Saharudin, Shahida, Abd Rahim, Nour El Huda, Halim, Mohd Dhiyaulhaq
Format: Article
Language:English
Published: IIUM Press 2022
Subjects:
Q Science (General)
QM Human anatomy
R Medicine (General)
RZ Other systems of medicine
Online Access:http://irep.iium.edu.my/99857/3/99857_The%20effect%20of%20prolonged%20fixation%20time%20on%20haematoxylin.pdf
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https://journals.iium.edu.my/ktn/index.php/ijohs/issue/view/9/8
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spelling my.iium.irep.998572022-09-09T08:43:45Z http://irep.iium.edu.my/99857/ The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue Mohd Radzuan, Hazulin Wan Omar, Wan Fatein Nabeila Saharudin, Shahida Abd Rahim, Nour El Huda Halim, Mohd Dhiyaulhaq Q Science (General) QM Human anatomy R Medicine (General) RZ Other systems of medicine The use of formalin in histopathological sample preparation is intended to preserve protein and cellular organelles1. It is argued that prolonged formalin fixation may lead to tissue shrinkage and hardening2,3. Reports on immunohistochemical studies showed that this does not cause reduction in antigen detection4. However, it is not known if prolonged formalin fixation can affect hematoxylin and eosin (H&E) staining quality. Our aim is to evaluate the effect of formalin in two different fixation times towards the quality of H&E staining adequacy in paraffin-embedded tissue blocks. Samples from Sprague-Dawley female rats, which include colon and placenta tissues, were harvested and processed to form formalin-fixed paraffin-embedded blocks. They were assessed in two durations: standard duration (SD) fixation up to 72 hours and prolonged duration (PD) up to 7 months in 10% neutral-buffered formalin (NBF). Thirty tissue sections from each study group were stained in modified Gill’s haematoxylin, and counter-stained in eosin before coverslipped with DPX. The slides were viewed with light microscope (Olympus BX51, Japan). We found that the staining quality was better among SD of placenta tissue as evidenced by basophilic appearance of basal spongiotrophoblast, and eosinophilic labyrinthine trophoblast. There was not much difference in terms of staining quality for colon, only that more artifacts can be observed among PD histological sections. In conclusion, prolonged fixation of colon and placental tissues in 10% NBF caused low-quality H&E staining. This could be attributed to diminished cellular organization and protein structure1. The difference between solid tissue and hollow organ sample also may contribute to the result. As much as we need to optimize sustainable resources, it should not compromise the quality of the outcome. We recommend preserving histopathological samples by adhering to the standard 72 hours duration of formalin-fixation, and archiving samples as paraffin-embedded tissue blocks for future studies. IIUM Press 2022 Article NonPeerReviewed application/pdf en http://irep.iium.edu.my/99857/3/99857_The%20effect%20of%20prolonged%20fixation%20time%20on%20haematoxylin.pdf Mohd Radzuan, Hazulin and Wan Omar, Wan Fatein Nabeila and Saharudin, Shahida and Abd Rahim, Nour El Huda and Halim, Mohd Dhiyaulhaq (2022) The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue. IIUM Journal of Orofacial and Health Sciences, 3 (Supplement 1). p. 52. E-ISSN 2735-0584 https://journals.iium.edu.my/ktn/index.php/ijohs/issue/view/9/8
institution Universiti Islam Antarabangsa Malaysia
building IIUM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider International Islamic University Malaysia
content_source IIUM Repository (IREP)
url_provider http://irep.iium.edu.my/
language English
topic Q Science (General)
QM Human anatomy
R Medicine (General)
RZ Other systems of medicine
spellingShingle Q Science (General)
QM Human anatomy
R Medicine (General)
RZ Other systems of medicine
Mohd Radzuan, Hazulin
Wan Omar, Wan Fatein Nabeila
Saharudin, Shahida
Abd Rahim, Nour El Huda
Halim, Mohd Dhiyaulhaq
The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue
description The use of formalin in histopathological sample preparation is intended to preserve protein and cellular organelles1. It is argued that prolonged formalin fixation may lead to tissue shrinkage and hardening2,3. Reports on immunohistochemical studies showed that this does not cause reduction in antigen detection4. However, it is not known if prolonged formalin fixation can affect hematoxylin and eosin (H&E) staining quality. Our aim is to evaluate the effect of formalin in two different fixation times towards the quality of H&E staining adequacy in paraffin-embedded tissue blocks. Samples from Sprague-Dawley female rats, which include colon and placenta tissues, were harvested and processed to form formalin-fixed paraffin-embedded blocks. They were assessed in two durations: standard duration (SD) fixation up to 72 hours and prolonged duration (PD) up to 7 months in 10% neutral-buffered formalin (NBF). Thirty tissue sections from each study group were stained in modified Gill’s haematoxylin, and counter-stained in eosin before coverslipped with DPX. The slides were viewed with light microscope (Olympus BX51, Japan). We found that the staining quality was better among SD of placenta tissue as evidenced by basophilic appearance of basal spongiotrophoblast, and eosinophilic labyrinthine trophoblast. There was not much difference in terms of staining quality for colon, only that more artifacts can be observed among PD histological sections. In conclusion, prolonged fixation of colon and placental tissues in 10% NBF caused low-quality H&E staining. This could be attributed to diminished cellular organization and protein structure1. The difference between solid tissue and hollow organ sample also may contribute to the result. As much as we need to optimize sustainable resources, it should not compromise the quality of the outcome. We recommend preserving histopathological samples by adhering to the standard 72 hours duration of formalin-fixation, and archiving samples as paraffin-embedded tissue blocks for future studies.
format Article
author Mohd Radzuan, Hazulin
Wan Omar, Wan Fatein Nabeila
Saharudin, Shahida
Abd Rahim, Nour El Huda
Halim, Mohd Dhiyaulhaq
author_facet Mohd Radzuan, Hazulin
Wan Omar, Wan Fatein Nabeila
Saharudin, Shahida
Abd Rahim, Nour El Huda
Halim, Mohd Dhiyaulhaq
author_sort Mohd Radzuan, Hazulin
title The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue
title_short The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue
title_full The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue
title_fullStr The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue
title_full_unstemmed The effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue
title_sort effect of prolonged fixation time on haematoxylin & eosin staining quality of rats colon and placenta tissue
publisher IIUM Press
publishDate 2022
url http://irep.iium.edu.my/99857/3/99857_The%20effect%20of%20prolonged%20fixation%20time%20on%20haematoxylin.pdf
http://irep.iium.edu.my/99857/
https://journals.iium.edu.my/ktn/index.php/ijohs/issue/view/9/8
_version_ 1744353530416726016
score 13.211869

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