PUBLICATION
TissUExM enables quantitative ultrastructural analysis in whole vertebrate embryos by expansion microscopy
- Authors
- Steib, E., Tetley, R., Laine, R.F., Norris, D.P., Mao, Y., Vermot, J.
- ID
- ZDB-PUB-221101-9
- Date
- 2022
- Source
- Cell reports methods 2: 100311 (Journal)
- Registered Authors
- Keywords
- centrioles, cilia, drosophila, expansion, mouse, super-resolution, whole embryos, zebrafish
- MeSH Terms
-
- Animals
- Drosophila
- Mice
- Microscopy*/methods
- Zebrafish*
- PubMed
- 36313808 Full text @ Cell Rep Methods
Citation
Steib, E., Tetley, R., Laine, R.F., Norris, D.P., Mao, Y., Vermot, J. (2022) TissUExM enables quantitative ultrastructural analysis in whole vertebrate embryos by expansion microscopy. Cell reports methods. 2:100311.
Abstract
Super-resolution microscopy reveals the molecular organization of biological structures down to the nanoscale. While it allows the study of protein complexes in single cells, small organisms, or thin tissue sections, there is currently no versatile approach for ultrastructural analysis compatible with whole vertebrate embryos. Here, we present tissue ultrastructure expansion microscopy (TissUExM), a method to expand millimeter-scale and mechanically heterogeneous whole embryonic tissues, including Drosophila wing discs, whole zebrafish, and mouse embryos. TissUExM is designed for the observation of endogenous proteins. It permits quantitative characterization of protein complexes in various organelles at super-resolution in a range of ∼3 mm-sized tissues using conventional microscopes. We demonstrate its strength by investigating tissue-specific ciliary architecture heterogeneity and ultrastructural defects observed upon ciliary protein overexpression. Overall, TissUExM is ideal for performing ultrastructural studies and molecular mapping in situ in whole embryos.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping