MICROSCOPY & IMAGING 75


Fig. 2. Comparison of deep tissue images of mouse cerebral neocortex sections with a 60x oil immersion (A) and a 60x silicone immersion (B) objective lens (Olympus UPLSAPO60XS2). Image courtesy of Motokazu Uchigashima & Masahiko Watanabe, Hokkaido University Graduate School of Medicine


mouse brain sections taken with 60x oil and 60x silicone immersion objectives. Bright fluorescence images were obtained with both sets of objectives, however X-Y images taken with silicone immersion objectives at a depth of 35µm were brighter with higher resolution (Fig.2). Te reduced refractive index mismatch of the silicone immersion objective enabled excellent performance in deep tissue observation on specimens rendered transparent.


Enabling 3D imaging Silicone immersion objectives are also proving vital in deep, high-contrast 3D live-cell time-lapse experiments. Tis is due to their reduced spherical aberrations, superior performance in maintaining focus for long periods of time, and resistance to changes in temperature. In a study at the Department of Genetic Engineering at Japan’s Kindai University, silicone objectives enabled sharp 3D fluorescence images of proteins,


DNA and other molecules in the zygote and individual cells during embryonic development. A 60x silicone immersion objective was used for 3D time-lapse imaging over four days during in vitro development of a mouse embryo from zygote to blastocyst.


Te team had been using an oil lens and water immersion objective, the previous standard for deep observation. Only by switching to a silicone immersion objective were the researchers able to view fluorescently labelled methylated DNA (mCherry-MBD-NLS) within the nuclei from the surface to the inner region – from the one cell zygote to the blastocyst stage (Fig.3).


Fig. 3. Live-cell imaging of a MethylRO embryo during pre-implantation development. Images acquired using Olympus silicone immersion objective UPLSAPO60XS. Image courtesy of Dr. Kazuo Yamagata, Kinki University


Precise fluorescence imaging Designed to have lower chromatic aberration, silicone objections are ideal for use with multiple fluorescent labels especially in colocalisation experiments. In a study at the RIKEN Brain Science Institute Laboratory for Cell Function Dynamics, researchers used a silicone immersion objective with an inverted


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