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Optimised tissue clearing minimises distortion and destruction during tissue delipidation

AIMS: A variety of tissue clearing techniques have been developed to render intact tissue transparent. For thicker samples, additional partial tissue delipidation is required before immersion into the final refractive index (RI)‐matching solution, which alone is often inadequate to achieve full tiss...

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Autores principales: Lee, Krit, Lai, Hei Ming, Soerensen, Maja Hoejvang, Hui, Edward Sai‐Kam, Ma, Victor Wan‐San, Cho, William Chi‐Shing, Ho, Yuen‐Shan, Chang, Raymond Chuen‐Chung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8048831/
https://www.ncbi.nlm.nih.gov/pubmed/33107057
http://dx.doi.org/10.1111/nan.12673
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author Lee, Krit
Lai, Hei Ming
Soerensen, Maja Hoejvang
Hui, Edward Sai‐Kam
Ma, Victor Wan‐San
Cho, William Chi‐Shing
Ho, Yuen‐Shan
Chang, Raymond Chuen‐Chung
author_facet Lee, Krit
Lai, Hei Ming
Soerensen, Maja Hoejvang
Hui, Edward Sai‐Kam
Ma, Victor Wan‐San
Cho, William Chi‐Shing
Ho, Yuen‐Shan
Chang, Raymond Chuen‐Chung
author_sort Lee, Krit
collection PubMed
description AIMS: A variety of tissue clearing techniques have been developed to render intact tissue transparent. For thicker samples, additional partial tissue delipidation is required before immersion into the final refractive index (RI)‐matching solution, which alone is often inadequate to achieve full tissue transparency. However, it is difficult to determine a sufficient degree of tissue delipidation, excess of which can result in tissue distortion and protein loss. Here, we aim to develop a clearing strategy that allows better monitoring and more precise determination of delipidation progress. METHODS: We combined the detergent sodium dodecyl sulphate (SDS) with OPTIClear, a RI‐matching solution, to form a strategy termed Accurate delipidation with Optimal Clearing (Accu‐OptiClearing). Accu‐OptiClearing allows for a better preview of the final tissue transparency achieved when immersed in OPTIClear alone just before imaging. We assessed for the changes in clearing rate, protein loss, degree of tissue distortion, and preservation of antigens. RESULTS: Partial delipidation using Accu‐OptiClearing accelerated tissue clearing and better preserved tissue structure and antigens than delipidation with SDS alone. Despite achieving similar transparency in the final OPTIClear solution, more lipids were retained in samples cleared with Accu‐OptiClearing compared to SDS. CONCLUSIONS: Combining the RI‐matching solution OPTIClear with detergents, Accu‐OptiClearing, can avoid excessive delipidation, leading to accelerated tissue clearing, less tissue damage and better preserved antigens.
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spelling pubmed-80488312021-04-20 Optimised tissue clearing minimises distortion and destruction during tissue delipidation Lee, Krit Lai, Hei Ming Soerensen, Maja Hoejvang Hui, Edward Sai‐Kam Ma, Victor Wan‐San Cho, William Chi‐Shing Ho, Yuen‐Shan Chang, Raymond Chuen‐Chung Neuropathol Appl Neurobiol Original Articles AIMS: A variety of tissue clearing techniques have been developed to render intact tissue transparent. For thicker samples, additional partial tissue delipidation is required before immersion into the final refractive index (RI)‐matching solution, which alone is often inadequate to achieve full tissue transparency. However, it is difficult to determine a sufficient degree of tissue delipidation, excess of which can result in tissue distortion and protein loss. Here, we aim to develop a clearing strategy that allows better monitoring and more precise determination of delipidation progress. METHODS: We combined the detergent sodium dodecyl sulphate (SDS) with OPTIClear, a RI‐matching solution, to form a strategy termed Accurate delipidation with Optimal Clearing (Accu‐OptiClearing). Accu‐OptiClearing allows for a better preview of the final tissue transparency achieved when immersed in OPTIClear alone just before imaging. We assessed for the changes in clearing rate, protein loss, degree of tissue distortion, and preservation of antigens. RESULTS: Partial delipidation using Accu‐OptiClearing accelerated tissue clearing and better preserved tissue structure and antigens than delipidation with SDS alone. Despite achieving similar transparency in the final OPTIClear solution, more lipids were retained in samples cleared with Accu‐OptiClearing compared to SDS. CONCLUSIONS: Combining the RI‐matching solution OPTIClear with detergents, Accu‐OptiClearing, can avoid excessive delipidation, leading to accelerated tissue clearing, less tissue damage and better preserved antigens. John Wiley and Sons Inc. 2020-12-02 2021-04 /pmc/articles/PMC8048831/ /pubmed/33107057 http://dx.doi.org/10.1111/nan.12673 Text en © 2020 The Authors. Neuropathology and Applied Neurobiology published by John Wiley & Sons Ltd on behalf of British Neuropathological Society https://creativecommons.org/licenses/by-nc/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited and is not used for commercial purposes.
spellingShingle Original Articles
Lee, Krit
Lai, Hei Ming
Soerensen, Maja Hoejvang
Hui, Edward Sai‐Kam
Ma, Victor Wan‐San
Cho, William Chi‐Shing
Ho, Yuen‐Shan
Chang, Raymond Chuen‐Chung
Optimised tissue clearing minimises distortion and destruction during tissue delipidation
title Optimised tissue clearing minimises distortion and destruction during tissue delipidation
title_full Optimised tissue clearing minimises distortion and destruction during tissue delipidation
title_fullStr Optimised tissue clearing minimises distortion and destruction during tissue delipidation
title_full_unstemmed Optimised tissue clearing minimises distortion and destruction during tissue delipidation
title_short Optimised tissue clearing minimises distortion and destruction during tissue delipidation
title_sort optimised tissue clearing minimises distortion and destruction during tissue delipidation
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8048831/
https://www.ncbi.nlm.nih.gov/pubmed/33107057
http://dx.doi.org/10.1111/nan.12673
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