Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy

TLDR

Large, living biological specimens pose challenges to optical imaging due to their absorptive and scattering properties. The authors developed selective plane illumination microscopy (SPIM) to generate multidimensional images of samples up to a few millimeters in size. SPIM combines two‑dimensional illumination with orthogonal camera detection to provide high‑resolution, optically sectioned imaging throughout millimeter‑scale samples, enabling rapid capture of transient biological phenomena with minimal photodamage. Using SPIM, the authors visualized all muscles in vivo in transgenic Medaka and captured embryogenesis in the relatively opaque Drosophila melanogaster.

Abstract

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.