Optical sectioning deep inside live embryos by selective plane illumination microscopy.
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.
Huisken
Jan
J
European Molecular Biology Laboratory (EMBL), Meyerhofstrasse 1, D-69117 Heidelberg, Germany. huisken@embl.de
Swoger
Jim
J
Del Bene
Filippo
F
Wittbrodt
Joachim
J
Stelzer
Ernst H K
EH
D019412:Anatomy, Cross-Sectional; D000818:Animals; D030801:Animals, Genetically Modified; D004331:Drosophila melanogaster / Q000196:embryology*; D004625:Embryo, Nonmammalian / Q000033:anatomy & histology* / Q000166:cytology; D047108:Embryonic Development; D049452:Green Fluorescent Proteins; D006321:Heart / Q000196:embryology*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D007834:Lasers; D008027:Light; D008164:Luminescent Proteins / Q000032:analysis; D008856:Microscopy, Fluorescence / Q000379:methods*; D009024:Morphogenesis; D009990:Oryzias / Q000196:embryology*
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins