26 April 2016 High-contrast 3D image acquisition using HiLo microscopy with an electrically tunable lens
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Abstract
We present a HiLo microscope with an electrically tunable lens for high-contrast three-dimensional image acquisition. HiLo microscopy combines wide field and speckled illumination images to create optically sectioned images. Additionally, the depth-of-field is not fixed, but can be adjusted between wide field and confocal-like axial resolution. We incorporate an electrically tunable lens in the HiLo microscope for axial scanning, to obtain three-dimensional data without the need of moving neither the sample nor the objective. The used adaptive lens consists of a transparent polydimethylsiloxane (PDMS) membrane into which an annular piezo bending actuator is embedded. A transparent fluid is filled between the membrane and the glass substrate. When actuated, the piezo generates a pressure in the lens which deflects the membrane and thus changes the refractive power. This technique enables a large tuning range of the refractive power between 1/f = (-24 . . . 25) 1/m. As the NA of the adaptive lens is only about 0.05, a fixed high-NA lens is included in the setup to provide high resolution. In this contribution, the scan properties and capabilities of the tunable lens in the HiLo microscope are analyzed. Eventually, exemplary measurements are presented and discussed.
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Katrin Philipp, André Smolarski, Andreas Fischer, Nektarios Koukourakis, Moritz Stürmer, Ulricke Wallrabe, Jürgen Czarske, "High-contrast 3D image acquisition using HiLo microscopy with an electrically tunable lens", Proc. SPIE 9890, Optical Micro- and Nanometrology VI, 98900A (26 April 2016); doi: 10.1117/12.2225596; https://doi.org/10.1117/12.2225596
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