Tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissue
Difficulty achieving complete, specific, and homogenous staining is a major bottleneck preventing the widespread use of tissue clearing techniques to image large volumes of human tissue. In this manuscript, we describe a procedure to rapidly design immunostaining protocols for antibody labeling of c...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2023-01-01
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| Series: | eLife |
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| Online Access: | https://elifesciences.org/articles/84112 |
| _version_ | 1811160387541794816 |
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| author | Theodore J Zwang Rachel E Bennett Maria Lysandrou Benjamin Woost Anqi Zhang Charles M Lieber Douglas S Richardson Bradley T Hyman |
| author_facet | Theodore J Zwang Rachel E Bennett Maria Lysandrou Benjamin Woost Anqi Zhang Charles M Lieber Douglas S Richardson Bradley T Hyman |
| author_sort | Theodore J Zwang |
| collection | DOAJ |
| description | Difficulty achieving complete, specific, and homogenous staining is a major bottleneck preventing the widespread use of tissue clearing techniques to image large volumes of human tissue. In this manuscript, we describe a procedure to rapidly design immunostaining protocols for antibody labeling of cleared brain tissue. We prepared libraries of 0.5–1.0 mm thick tissue sections that are fixed, pre-treated, and cleared via similar, but different procedures to optimize staining conditions for a panel of antibodies. Results from a library of mouse tissue correlate well with results from a similarly prepared library of human brain tissue, suggesting mouse tissue is an adequate substitute for protocol optimization. These data show that procedural differences do not influence every antibody-antigen pair in the same way, and minor changes can have deleterious effects, therefore, optimization should be conducted for each target. The approach outlined here will help guide researchers to successfully label a variety of targets, thus removing a major hurdle to accessing the rich 3D information available in large, cleared human tissue volumes. |
| first_indexed | 2024-04-10T05:56:30Z |
| format | Article |
| id | doaj.art-bac94001b40d422c9fd51c43ea689c90 |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-10T05:56:30Z |
| publishDate | 2023-01-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-bac94001b40d422c9fd51c43ea689c902023-03-03T15:01:09ZengeLife Sciences Publications LtdeLife2050-084X2023-01-011210.7554/eLife.84112Tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissueTheodore J Zwang0https://orcid.org/0000-0002-6815-4482Rachel E Bennett1Maria Lysandrou2Benjamin Woost3Anqi Zhang4https://orcid.org/0000-0001-6121-8095Charles M Lieber5Douglas S Richardson6https://orcid.org/0000-0002-3189-2190Bradley T Hyman7https://orcid.org/0000-0002-7959-9401Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, United States; Harvard Medical School, Boston, United States; Massachusetts Alzheimer’s Disease Research Center, Charlestown, United StatesDepartment of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, United States; Harvard Medical School, Boston, United States; Massachusetts Alzheimer’s Disease Research Center, Charlestown, United StatesDepartment of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, United StatesDepartment of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, United StatesDepartment of Chemical Engineering, Stanford University, Stanford, United StatesDepartment of Chemistry and Chemical Biology, Harvard University, Cambridge, United StatesDepartment of Molecular and Cellular Biology and Harvard Center for Biological Imaging, Harvard University, Cambridge, United StatesDepartment of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, United States; Harvard Medical School, Boston, United States; Massachusetts Alzheimer’s Disease Research Center, Charlestown, United StatesDifficulty achieving complete, specific, and homogenous staining is a major bottleneck preventing the widespread use of tissue clearing techniques to image large volumes of human tissue. In this manuscript, we describe a procedure to rapidly design immunostaining protocols for antibody labeling of cleared brain tissue. We prepared libraries of 0.5–1.0 mm thick tissue sections that are fixed, pre-treated, and cleared via similar, but different procedures to optimize staining conditions for a panel of antibodies. Results from a library of mouse tissue correlate well with results from a similarly prepared library of human brain tissue, suggesting mouse tissue is an adequate substitute for protocol optimization. These data show that procedural differences do not influence every antibody-antigen pair in the same way, and minor changes can have deleterious effects, therefore, optimization should be conducted for each target. The approach outlined here will help guide researchers to successfully label a variety of targets, thus removing a major hurdle to accessing the rich 3D information available in large, cleared human tissue volumes.https://elifesciences.org/articles/84112cell biologyneurosciencehumanmouse |
| spellingShingle | Theodore J Zwang Rachel E Bennett Maria Lysandrou Benjamin Woost Anqi Zhang Charles M Lieber Douglas S Richardson Bradley T Hyman Tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissue eLife cell biology neuroscience human mouse |
| title | Tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissue |
| title_full | Tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissue |
| title_fullStr | Tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissue |
| title_full_unstemmed | Tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissue |
| title_short | Tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissue |
| title_sort | tissue libraries enable rapid determination of conditions that preserve antibody labeling in cleared mouse and human tissue |
| topic | cell biology neuroscience human mouse |
| url | https://elifesciences.org/articles/84112 |
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