76 MICROSCOPY & IMAGING
Fig. 5. Merged image of placenta cells showing precise colocalisation of markers Image courtesy of Asako Sakaue-Sawano & Atsushi
Miyawaki, RIKEN Brain Science Institute Laboratory for Cell Function Dynamics
Fig. 4. Observation of cells in a whole mouse placenta (sequential tomography) using a 40x silicone immersion objective (Olympus UPLSAPO40XS) Image courtesy of Asako Sakaue-Sawano RIKEN Brain Science Institute Laboratory for Cell Function Dynamics
confocal laser-scanning microscope to make high-resolution deep tissue observations of morphologically intact cells within a whole mouse placenta without needing to section it. It was possible to precisely observe colocalisation of
ubiquitination-based cell cycle indicators (Fucci), (S/G2/M-Green, G1-Red) and the nucleus (DAPI-Blue) (Figs 4 and 5).
Breaking new ground Silicone objectives are even driving
new technologies in live-cell imaging. Te world’s first long-term, real-time observation of plant zygote embryogenesis was made possible by combining a new ovule culture system with a microscope capable of high sensitivity embryo imaging inside the ovule and covered by multiple layers of cells. In this study by Te Optical
Technology Group of ERATO Higashiyama Live-Holonics Project at Japan’s Nagoya University, long-term live-cell imaging of plant zygote division and growth was made possible by the development of a special medium and a new microdevice, the use of which included a silicone immersion objective (Olympus UPLSAPO30XS) (Fig.6). Tis objective has a high numerical aperture of 1.05 and a long working distance of 0.8mm, which is required for deep, high- definition imaging while retaining a wide field of view. With this set-up the process of embryogenesis was stably observed in real time for 67 hours from the early to late embryo.
Summary As shown by the examples discussed here, silicone immersion objectives are proving to be the perfect partner for live-cell imaging applications, delivering unmatched resolution, clarity and reproducibility.
Fig. 6. Microdevice with a 30x silicone immersion objective (Olympus UPLSAPO30XS). Multipoint time-lapse images were acquired with a motorised XY stage
www.scientistlive.com
Dr Jan Barghaan is with Olympus Europa.
www.olympus-europa.com
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