Broad applicability of a streamlined Ethyl Cinnamate-based clearing procedure
| Autor: | Jürgen A. Knoblich, François Bonnay, Yuka Taniguchi, Katharina Meixner, Pawel Pasierbek, Daniel Reumann, Prayag Murawala, Elly M. Tanaka, Wouter Masselink |
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| Rok vydání: | 2019 |
| Předmět: |
Biology
Article Xenopus laevis 03 medical and health sciences chemistry.chemical_compound Imaging Three-Dimensional 0302 clinical medicine Clearing medicine Animals Humans Brainbow Molecular Biology Zebrafish Fluorescent Dyes 030304 developmental biology 0303 health sciences Tissue clearing Ethyl cinnamate Ambystoma mexicanum Organoids Drosophila melanogaster medicine.anatomical_structure Microscopy Fluorescence chemistry Cinnamates mCherry Biological system 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Development |
| ISSN: | 1477-9129 0950-1991 |
| DOI: | 10.1242/dev.166884 |
| Popis: | Turbidity and opaqueness are inherent properties of tissues that limit the capacity to acquire microscopic images through large tissues. Creating a uniform refractive index, known as tissue clearing, overcomes most of these issues. These methods have enabled researchers to image large and complex 3D structures with unprecedented depth and resolution. However, tissue clearing has been adopted to a limited extent due to a combination of cost, time, complexity of existing methods and potential negative impact on fluorescence signal. Here, we describe 2Eci (2nd generation ethyl cinnamate-based clearing), which can be used to clear a wide range of tissues in several species, including human organoids, Drosophila melanogaster, zebrafish, axolotl and Xenopus laevis, in as little as 1-5 days, while preserving a broad range of fluorescent proteins, including GFP, mCherry, Brainbow and Alexa-conjugated fluorophores. Ethyl cinnamate is non-toxic and can easily be used in multi-user microscope facilities. This method opens up tissue clearing to a much broader group of researchers due to its ease of use, the non-toxic nature of ethyl cinnamate and broad applicability. |
| Databáze: | OpenAIRE |
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