| Autor: |
Subiran Adrados, Cristina, Yu, Qinghao, Bolaños Castro, Lizbeth Airais, Rodriguez Cabrera, Luis Alberto, Yun, Maximina Hee |
| Předmět: |
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| Zdroj: |
Developmental Dynamics; Jun2021, Vol. 250 Issue 6, p902-915, 14p |
| Abstrakt: |
Background: Salamander limb regeneration is a complex biological process that entails the orchestration of multiple cellular and molecular mechanisms in a three‐dimensional space. Hence, a comprehensive understanding of this process requires whole‐structure level explorations. Recent advances in imaging and optical clearing methods have transformed the study of regenerative phenomena, allowing the three‐dimensional visualization of structures and entire organisms. Results: Here we introduce Salamander‐Eci, a rapid and robust optical clearing protocol optimized for the widely used axolotl model, which allows simultaneous immunohistochemistry and Click‐chemistry detection with minimal volume disruption. We provide examples of its application, from whole larva to adult limbs and organs, and complement it with an image analysis pipeline for volumetric cell quantification. Further, we offer a detailed 3D quantitation of cell proliferation throughout axolotl limb regeneration. Conclusions: Salamander‐Eci enables the comprehensive volumetric analysis of regenerative phenomena at both local and systemic levels. Key Findings: Salamander‐Eci, a rapid and robust optical clearing protocol with minimal tissue disruption optimised for the axolotl modelSimultaneous immunohistochemistry and Click chemistry detectionVolumetric quantification pipeline3D quantitation of cell proliferation throughout axolotl limb regeneration [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
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