Simultaneous multiple-level magnification selective plane illumination microscopy (sMx-SPIM) imaging system

Abstract

We report an optical-based microscopy imaging technology—simultaneous multiple-level magnification selective plane illumination microscopy (sMx-SPIM) imaging system—that addresses a longstanding technological challenge of obtaining images, specifically of biological specimen non-destructively, at different field of views (FOVs) and spatial resolutions (or magnification powers) simultaneously in real-time. This imaging system provides not only 3D images but also time-resolved sequential images with temporal resolution ∼ msec. Magnification powers (or FOVs) of the individual images can be controlled independently that is achieved by housing two separate detection arms, in SPIM imaging system, fitted with objective lenses of different magnification powers. This unique feature holds promises to observe and study: (a) sub-microscopic details and entire structure of biological specimen side-by-side simultaneously and (b) spatio-temporal dynamics of functional activities of biological specimen. For validation study of robustness of the proposed sMx-SPIM imaging system, experiments are conducted in various biological samples including Danio rerio (zebrafish) embryo, Drosophila melanogaster, Allium cepa root, and A549 cell line. Experimental results demonstrate that the study is of significant impacts from two aspects, viz., technological implication and biological applications.