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Digital Fly Embryo

CEF Investigator Ernst Stelzer and colleagues from the European Molecular Biology Laboratory in Heidelberg, the University of Heidelberg and the Sloan-Kettering Institute in New York have created the Fly Digital Embryo. In work reported on 4 July 2010 in the journal Nature Methods, they were able to film the development of fruit flys as well as that the eyes and midbrain of zebra fish.

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. Ernst Stelzer and his team combined digital scanned laser light-sheet fluorescence microscopy with incoherent structured-illumination microscopy (DSLM-SI) and created structured-illumination patterns with continuously adjustable frequencies. The improved 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. The scientists conducted imaging of zebrafish development for 58 h and fast multiple-view imaging of early Drosophila melanogaster development. They reconstructed cell positions over time from the Drosophila DSLM-SI data and created a fly digital embryo. Their improvement in technique will help to shed light on processes and organisms, which have so far been under-studied because they could not be studied well under a microscope. All data, images and videos are freely available online, alongside the data from the digital embryo, at www.digital-embryo.org.

Full reference: Philipp J. Keller, Annette D. Schmidt, Anthony Santella, Khaled Khairy, Zhirong Bao, Joachim Wittbrodt and Ernst H.K. Stelzer. 2010. Fast, high-contrast imaging of animal development by scanned light sheet-based structured illumination microscopy. Nature Methods, 4 July 2010 (Advance Online Publication, DOI 10.1038/nmeth.1476). Link