Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy Journal Article

   

Authors:	Keller, P. J.; Schmidt, A. D.; Santella, A.; Khairy, K.; Bao, Z.; Wittbrodt, J.; Stelzer, E. H. K.
Article Title:	Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy
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. © 2010 Nature America, Inc. All rights reserved.
Keywords:	microscopy; nonhuman; animals; embryo development; animalia; embryo, nonmammalian; image quality; imaging; drosophila melanogaster; developmental stage; zebra fish; zebrafish; danio rerio; insect development; illumination
Journal Title:	Nature Methods
Volume:	7
Issue:	8
ISSN:	1548-7091
Publisher:	Nature Publishing Group
Date Published:	2010-08-01
Start Page:	637
End Page:	642
Language:	English
DOI:	10.1038/nmeth.1476
PUBMED:	20601950
PROVIDER:	scopus
PMCID:	PMC4418465
DOI/URL:	http://www.scopus.com/inward/record.url?eid=2-s2.0-77955172060&partnerID=40&md5=a18bb885b0d2e1c02859b0e217752fc3
Notes:	--- - "Cited By (since 1996): 4" - "Export Date: 20 April 2011" - "Source: Scopus"

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