See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates
Studies of morphology and developmental patterning in adult stages of many invertebrates are hindered by opaque structures, such as shells, skeletal elements, and pigment granules that block or refract light and necessitate sectioning for observation of internal features. An inherent challenge in st...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Springer Science and Business Media LLC
2024
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| Online Access: | https://hdl.handle.net/1721.1/155668 |
| _version_ | 1824458102596960256 |
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| author | Clarke, D. N. Formery, L. Lowe, C. J. |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Clarke, D. N. Formery, L. Lowe, C. J. |
| author_sort | Clarke, D. N. |
| collection | MIT |
| description | Studies of morphology and developmental patterning in adult stages of many invertebrates are hindered by opaque structures, such as shells, skeletal elements, and pigment granules that block or refract light and necessitate sectioning for observation of internal features. An inherent challenge in studies relying on surgical approaches is that cutting tissue is semi-destructive, and delicate structures, such as axonal processes within neural networks, are computationally challenging to reconstruct once disrupted. To address this problem, we developed See-Star, a hydrogel-based tissue clearing protocol to render the bodies of opaque and calcified invertebrates optically transparent while preserving their anatomy in an unperturbed state, facilitating molecular labeling and observation of intact organ systems. The resulting protocol can clear large (> 1 cm3) specimens to enable deep-tissue imaging, and is compatible with molecular techniques, such as immunohistochemistry and in situ hybridization to visualize protein and mRNA localization. To test the utility of this method, we performed a whole-mount imaging study of intact nervous systems in juvenile echinoderms and molluscs and demonstrate that See-Star allows for comparative studies to be extended far into development, facilitating insights into the anatomy of juveniles and adults that are usually not amenable to whole-mount imaging. |
| first_indexed | 2024-09-23T11:43:30Z |
| format | Article |
| id | mit-1721.1/155668 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2025-02-19T04:20:33Z |
| publishDate | 2024 |
| publisher | Springer Science and Business Media LLC |
| record_format | dspace |
| spelling | mit-1721.1/1556682025-01-09T04:52:41Z See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates Clarke, D. N. Formery, L. Lowe, C. J. Massachusetts Institute of Technology. Department of Biology Studies of morphology and developmental patterning in adult stages of many invertebrates are hindered by opaque structures, such as shells, skeletal elements, and pigment granules that block or refract light and necessitate sectioning for observation of internal features. An inherent challenge in studies relying on surgical approaches is that cutting tissue is semi-destructive, and delicate structures, such as axonal processes within neural networks, are computationally challenging to reconstruct once disrupted. To address this problem, we developed See-Star, a hydrogel-based tissue clearing protocol to render the bodies of opaque and calcified invertebrates optically transparent while preserving their anatomy in an unperturbed state, facilitating molecular labeling and observation of intact organ systems. The resulting protocol can clear large (> 1 cm3) specimens to enable deep-tissue imaging, and is compatible with molecular techniques, such as immunohistochemistry and in situ hybridization to visualize protein and mRNA localization. To test the utility of this method, we performed a whole-mount imaging study of intact nervous systems in juvenile echinoderms and molluscs and demonstrate that See-Star allows for comparative studies to be extended far into development, facilitating insights into the anatomy of juveniles and adults that are usually not amenable to whole-mount imaging. 2024-07-12T15:47:11Z 2024-07-12T15:47:11Z 2024-06-25 2024-06-30T03:11:20Z Article http://purl.org/eprint/type/JournalArticle 2041-9139 https://hdl.handle.net/1721.1/155668 Clarke, D.N., Formery, L. & Lowe, C.J. See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates. EvoDevo 15, 8 (2024). PUBLISHER_CC en 10.1186/s13227-024-00228-0 EvoDevo Creative Commons Attribution https://creativecommons.org/licenses/by/4.0/ The Author(s) application/pdf Springer Science and Business Media LLC BioMed Central |
| spellingShingle | Clarke, D. N. Formery, L. Lowe, C. J. See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates |
| title | See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates |
| title_full | See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates |
| title_fullStr | See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates |
| title_full_unstemmed | See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates |
| title_short | See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates |
| title_sort | see star a versatile hydrogel based protocol for clearing large opaque and calcified marine invertebrates |
| url | https://hdl.handle.net/1721.1/155668 |
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