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...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Academic Press Inc.
2024
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| Online Access: | http://psasir.upm.edu.my/id/eprint/115064/ http://psasir.upm.edu.my/id/eprint/115064/1/115064.pdf |
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| 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 |
| 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 |
| building | UPM Institutional Repository |
| collection | Online Access |
| 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. |
| first_indexed | 2025-11-15T14:24:22Z |
| format | Article |
| id | upm-115064 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T14:24:22Z |
| publishDate | 2024 |
| publisher | Academic Press Inc. |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-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 |
| 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 |
| 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_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_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_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 |
| url | http://psasir.upm.edu.my/id/eprint/115064/ http://psasir.upm.edu.my/id/eprint/115064/ http://psasir.upm.edu.my/id/eprint/115064/ http://psasir.upm.edu.my/id/eprint/115064/1/115064.pdf |