EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application

Coolant-assisted liquid nitrogen (LN) flash freezing of frozen tissues has been widely adopted to preserve tissue morphology for histopathological annotations in mass spectrometry-based spatial proteomics techniques. However, existing coolants pose health risks upon inhalation and are expensive. To...

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Main Authors: Tan, Yong Chiang, Mustangin, Muaatamarulain, Rosli, Nurwahyuna, Wan Ahmad Kammal, Wan Syahira Ellani, Md Isa, Nurismah, Low, Teck Yew, Lee, Pey Yee, Chellappan, Dinesh Kumar, Jarmin, Razman, Zuhdi, Zamri, Azman, Azlanudin, Ian, Chik, Mohamed Yusof, Nursuhadah, Lim, Lay Cheng
Format: Article
Language:English
Published: Academic Press Inc. 2024
Online Access:http://psasir.upm.edu.my/id/eprint/115064/1/115064.pdf
http://psasir.upm.edu.my/id/eprint/115064/
https://linkinghub.elsevier.com/retrieve/pii/S0011224023003425
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Institution: Universiti Putra Malaysia
Language: English
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spelling my.upm.eprints.1150642025-02-18T09:17:16Z http://psasir.upm.edu.my/id/eprint/115064/ EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application Tan, Yong Chiang Mustangin, Muaatamarulain Rosli, Nurwahyuna Wan Ahmad Kammal, Wan Syahira Ellani Md Isa, Nurismah Low, Teck Yew Lee, Pey Yee Chellappan, Dinesh Kumar Jarmin, Razman Zuhdi, Zamri Azman, Azlanudin Ian, Chik Mohamed Yusof, Nursuhadah Lim, Lay Cheng Coolant-assisted liquid nitrogen (LN) flash freezing of frozen tissues has been widely adopted to preserve tissue morphology for histopathological annotations in mass spectrometry-based spatial proteomics techniques. However, existing coolants pose health risks upon inhalation and are expensive. To overcome this challenge, we present our pilot study by introducing the EtOH-LN workflow, which demonstrates the feasibility of using 95 % ethanol as a safer and easily accessible alternative to existing coolants for LN-based cryoembedding of frozen tissues. Our study reveals that both the EtOH-LN and LN-only cryoembedding workflows exhibit significantly reduced freezing artifacts compared to cryoembedding in cryostat (p < 0.005), while EtOH-LN (SD = 0.56) generates more consistent results compared to LN-only (SD = 1.29). We have modified a previously reported morphology restoration method to incorporate the EtOH-LN workflow, which successfully restored the tissue architecture from freezing artifacts (p < 0.05). Additional studies are required to validate the impact of the EtOH-LN workflow on the molecular profiles of tissues. Academic Press Inc. 2024 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/115064/1/115064.pdf Tan, Yong Chiang and Mustangin, Muaatamarulain and Rosli, Nurwahyuna and Wan Ahmad Kammal, Wan Syahira Ellani and Md Isa, Nurismah and Low, Teck Yew and Lee, Pey Yee and Chellappan, Dinesh Kumar and Jarmin, Razman and Zuhdi, Zamri and Azman, Azlanudin and Ian, Chik and Mohamed Yusof, Nursuhadah and Lim, Lay Cheng (2024) EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application. Cryobiology, 114. art. no. 104843. ISSN 0011-2240; eISSN: 1090-2392 https://linkinghub.elsevier.com/retrieve/pii/S0011224023003425 10.1016/j.cryobiol.2023.104843
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description Coolant-assisted liquid nitrogen (LN) flash freezing of frozen tissues has been widely adopted to preserve tissue morphology for histopathological annotations in mass spectrometry-based spatial proteomics techniques. However, existing coolants pose health risks upon inhalation and are expensive. To overcome this challenge, we present our pilot study by introducing the EtOH-LN workflow, which demonstrates the feasibility of using 95 % ethanol as a safer and easily accessible alternative to existing coolants for LN-based cryoembedding of frozen tissues. Our study reveals that both the EtOH-LN and LN-only cryoembedding workflows exhibit significantly reduced freezing artifacts compared to cryoembedding in cryostat (p < 0.005), while EtOH-LN (SD = 0.56) generates more consistent results compared to LN-only (SD = 1.29). We have modified a previously reported morphology restoration method to incorporate the EtOH-LN workflow, which successfully restored the tissue architecture from freezing artifacts (p < 0.05). Additional studies are required to validate the impact of the EtOH-LN workflow on the molecular profiles of tissues.
format Article
author Tan, Yong Chiang
Mustangin, Muaatamarulain
Rosli, Nurwahyuna
Wan Ahmad Kammal, Wan Syahira Ellani
Md Isa, Nurismah
Low, Teck Yew
Lee, Pey Yee
Chellappan, Dinesh Kumar
Jarmin, Razman
Zuhdi, Zamri
Azman, Azlanudin
Ian, Chik
Mohamed Yusof, Nursuhadah
Lim, Lay Cheng
spellingShingle Tan, Yong Chiang
Mustangin, Muaatamarulain
Rosli, Nurwahyuna
Wan Ahmad Kammal, Wan Syahira Ellani
Md Isa, Nurismah
Low, Teck Yew
Lee, Pey Yee
Chellappan, Dinesh Kumar
Jarmin, Razman
Zuhdi, Zamri
Azman, Azlanudin
Ian, Chik
Mohamed Yusof, Nursuhadah
Lim, Lay Cheng
EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application
author_facet Tan, Yong Chiang
Mustangin, Muaatamarulain
Rosli, Nurwahyuna
Wan Ahmad Kammal, Wan Syahira Ellani
Md Isa, Nurismah
Low, Teck Yew
Lee, Pey Yee
Chellappan, Dinesh Kumar
Jarmin, Razman
Zuhdi, Zamri
Azman, Azlanudin
Ian, Chik
Mohamed Yusof, Nursuhadah
Lim, Lay Cheng
author_sort Tan, Yong Chiang
title EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application
title_short EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application
title_full EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application
title_fullStr EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application
title_full_unstemmed EtOH-LN cryoembedding workflow to minimize freezing artifact in frozen tissues: A pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application
title_sort etoh-ln cryoembedding workflow to minimize freezing artifact in frozen tissues: a pilot study in preparing tissues compatible with mass spectrometry-based spatial proteomics application
publisher Academic Press Inc.
publishDate 2024
url http://psasir.upm.edu.my/id/eprint/115064/1/115064.pdf
http://psasir.upm.edu.my/id/eprint/115064/
https://linkinghub.elsevier.com/retrieve/pii/S0011224023003425
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