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Science 13 August 2004:
Vol. 305. no. 5686, pp. 1007 - 1009
DOI: 10.1126/science.1100035
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Reports
Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy
Jan Huisken,*
Jim Swoger,
Filippo Del Bene,
Joachim Wittbrodt,
Ernst H. K. Stelzer*
Large, living biological specimens present challenges to existing optical imaging techniques because of their absorptive and scattering properties. We developed selective plane illumination microscopy (SPIM) to generate multidimensional images of samples up to a few millimeters in size. The system combines two-dimensional illumination with orthogonal camera-based detection to achieve high-resolution, optically sectioned imaging throughout the sample, with minimal photodamage and at speeds capable of capturing transient biological phenomena. We used SPIM to visualize all muscles in vivo in the transgenic Medaka line Arnie, which expresses green fluorescent protein in muscle tissue. We also demonstrate that SPIM can be applied to visualize the embryogenesis of the relatively opaque Drosophila melanogaster in vivo.
European Molecular Biology Laboratory (EMBL), Meyerhofstraße 1, D-69117 Heidelberg, Germany.
* To whom correspondence should be addressed. E-mail: huisken{at}embl.de (J.H.) and stelzer{at}embl.de (E.H.K.S.)
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