Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy
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 requi...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article (Journal) |
| Language: | English |
| Published: |
04 July 2010
|
| In: |
Nature methods
Year: 2010, Volume: 7, Issue: 8, Pages: 637-642 |
| ISSN: | 1548-7105 |
| DOI: | 10.1038/nmeth.1476 |
| Online Access: | Verlag, Volltext: http://dx.doi.org/10.1038/nmeth.1476 Verlag, Volltext: https://www.nature.com/nmeth/journal/v7/n8/full/nmeth.1476.html 
|
| Author Notes: | Philipp J. Keller, Annette D. Schmidt, Anthony Santella, Khaled Khairy, Zhirong Bao, Joachim Wittbrodt & Ernst H. K. Stelzer |
| Summary: | 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. |
|---|---|
| Item Description: | Gesehen am 02.06.2017 |
| Physical Description: | Online Resource |
| ISSN: | 1548-7105 |
| DOI: | 10.1038/nmeth.1476 |