Optical sectioning deep inside live embryos by selective plane illumination microscopy

J Huisken, J Swoger, F Del Bene, J Wittbrodt… - Science, 2004 - science.org
Science, 2004science.org
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 …
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.
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