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61 Janelia Publications

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    Keller Lab
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    08/01/10 | Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy.
    Keller PJ, Schmidt AD, Santella A, Khairy K, Zhirong Bao , Wittbrodt J, Stelzer EH
    Nature Methods. 08/2010;7(8):637-42. doi: 10.1038/nmeth.1476

    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.

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