PUBLICATION
Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy
- Authors
- Keller, P.J., Schmidt, A.D., Santella, A., Khairy, K., Bao, Z., Wittbrodt, J., and Stelzer, E.H.
- ID
- ZDB-PUB-100719-16
- Date
- 2010
- Source
- Nature Methods 7(8): 637-642 (Journal)
- Registered Authors
- Keller, Philipp, Schmidt, Anne
- Keywords
- none
- MeSH Terms
-
- Animals
- Drosophila melanogaster/growth & development
- Embryo, Nonmammalian
- Microscopy/instrumentation*
- Microscopy/methods*
- Zebrafish/growth & development
- PubMed
- 20601950 Full text @ Nat. Methods
Citation
Keller, P.J., Schmidt, A.D., Santella, A., Khairy, K., Bao, Z., Wittbrodt, J., and Stelzer, E.H. (2010) Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy. Nature Methods. 7(8):637-642.
Abstract
Recording light-microscopy images of large, nontransparent specimens, such as developing multicellular organisms, is complicated by decreased contrast resulting from light scattering. Early zebrafish development can be captured by standard light-sheet microscopy, but new imaging strategies are required to obtain high-quality data of late development or of less transparent organisms. We combined digital scanned laser light-sheet fluorescence microscopy with incoherent structured-illumination microscopy (DSLM-SI) and created structured-illumination patterns with continuously adjustable frequencies. Our method discriminates the specimen-related scattered background from signal fluorescence, thereby removing out-of-focus light and optimizing the contrast of in-focus structures. DSLM-SI provides rapid control of the illumination pattern, exceptional imaging quality and high imaging speeds. We performed long-term imaging of zebrafish development for 58 h and fast multiple-view imaging of early Drosophila melanogaster development. We reconstructed cell positions over time from the Drosophila DSLM-SI data and created a fly digital embryo.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping