Comparison of tissue processing methods for microvascular visualization in axolotls
The vascular system, the pipeline for oxygen and nutrient delivery to tissues, is essential for vertebrate development, growth, injury repair, and regeneration. With their capacity to regenerate entire appendages throughout their lifespan, axolotls are an unparalleled model for vertebrate regenerati...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2017-01-01
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| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016117300262 |
| _version_ | 1818515166652792832 |
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| author | Rodrigo Montoro Renee Dickie |
| author_facet | Rodrigo Montoro Renee Dickie |
| author_sort | Rodrigo Montoro |
| collection | DOAJ |
| description | The vascular system, the pipeline for oxygen and nutrient delivery to tissues, is essential for vertebrate development, growth, injury repair, and regeneration. With their capacity to regenerate entire appendages throughout their lifespan, axolotls are an unparalleled model for vertebrate regeneration, but they lack many of the molecular tools that facilitate vascular imaging in other animal models. The determination of vascular metrics requires high quality image data for the discrimination of vessels from background tissue. Quantification of the vasculature using perfused, cleared specimens is well-established in mammalian systems, but has not been widely employed in amphibians. The objective of this study was to optimize tissue preparation methods for the visualization of the microvascular network in axolotls, providing a basis for the quantification of regenerative angiogenesis. To accomplish this aim, we performed intracardiac perfusion of pigment-based contrast agents and evaluated aqueous and non-aqueous clearing techniques. The methods were verified by comparing the quality of the vascular images and the observable vascular density across treatment groups. Simple and inexpensive, these tissue processing techniques will be of use in studies assessing vascular growth and remodeling within the context of regeneration. Advantages of this method include:
• Higher contrast of the vasculature within the 3D context of the surrounding tissue
• Enhanced detection of microvasculature facilitating vascular quantification
• Compatibility with other labeling techniques |
| first_indexed | 2024-12-11T00:25:25Z |
| format | Article |
| id | doaj.art-ca3225e2de854592a2a7840ece8acc13 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-11T00:25:25Z |
| publishDate | 2017-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-ca3225e2de854592a2a7840ece8acc132022-12-22T01:27:34ZengElsevierMethodsX2215-01612017-01-014C26527310.1016/j.mex.2017.08.001Comparison of tissue processing methods for microvascular visualization in axolotlsRodrigo MontoroRenee DickieThe vascular system, the pipeline for oxygen and nutrient delivery to tissues, is essential for vertebrate development, growth, injury repair, and regeneration. With their capacity to regenerate entire appendages throughout their lifespan, axolotls are an unparalleled model for vertebrate regeneration, but they lack many of the molecular tools that facilitate vascular imaging in other animal models. The determination of vascular metrics requires high quality image data for the discrimination of vessels from background tissue. Quantification of the vasculature using perfused, cleared specimens is well-established in mammalian systems, but has not been widely employed in amphibians. The objective of this study was to optimize tissue preparation methods for the visualization of the microvascular network in axolotls, providing a basis for the quantification of regenerative angiogenesis. To accomplish this aim, we performed intracardiac perfusion of pigment-based contrast agents and evaluated aqueous and non-aqueous clearing techniques. The methods were verified by comparing the quality of the vascular images and the observable vascular density across treatment groups. Simple and inexpensive, these tissue processing techniques will be of use in studies assessing vascular growth and remodeling within the context of regeneration. Advantages of this method include: • Higher contrast of the vasculature within the 3D context of the surrounding tissue • Enhanced detection of microvasculature facilitating vascular quantification • Compatibility with other labeling techniqueshttp://www.sciencedirect.com/science/article/pii/S2215016117300262Microvascular visualization in axolotls |
| spellingShingle | Rodrigo Montoro Renee Dickie Comparison of tissue processing methods for microvascular visualization in axolotls MethodsX Microvascular visualization in axolotls |
| title | Comparison of tissue processing methods for microvascular visualization in axolotls |
| title_full | Comparison of tissue processing methods for microvascular visualization in axolotls |
| title_fullStr | Comparison of tissue processing methods for microvascular visualization in axolotls |
| title_full_unstemmed | Comparison of tissue processing methods for microvascular visualization in axolotls |
| title_short | Comparison of tissue processing methods for microvascular visualization in axolotls |
| title_sort | comparison of tissue processing methods for microvascular visualization in axolotls |
| topic | Microvascular visualization in axolotls |
| url | http://www.sciencedirect.com/science/article/pii/S2215016117300262 |
| work_keys_str_mv | AT rodrigomontoro comparisonoftissueprocessingmethodsformicrovascularvisualizationinaxolotls AT reneedickie comparisonoftissueprocessingmethodsformicrovascularvisualizationinaxolotls |