Axial scanning and spherical aberration correction in confocal microscopy employing an adaptive lens

We present a fluid-membrane lens with two piezoelectric actuators that offer versatile, circular symmetric lens surface shaping. A wavefront-measurement-based control system ensures robustness against creeping and hysteresis effects of the piezoelectric actuators. We apply the adaptive lens to correct synthetic aberrations induced by a deformable mirror. The results suggest that the lens is able to correct spherical aberrations with standard Zernike coefficients between 0 μm and 1 μm, while operating at refractive powers up to about 4m-1. We apply the adaptive lens in a custom-built confocal microscope to allow simultaneous axial scanning and spherical aberration tuning. The confocal microscope is extended by an additional phase measurement system to include the control algorithm. To verify our approach, we use the maximum intensity and the axial FWHM of the overall confocal point spread function as figures of merit. We further discuss the ability of the adaptive lens to correct specimen-induced aberrations in a confocal microscope.

[1]  Ulrike Wallrabe,et al.  A Compact, Large-Aperture Tunable Lens with Adaptive Spherical Correction , 2014, 2014 International Symposium on Optomechatronic Technologies.

[2]  Florian Lemke,et al.  Adaptive lenses for axial scanning in HiLo microscopy , 2017 .

[3]  Jürgen Czarske,et al.  High-contrast 3D image acquisition using HiLo microscopy with an electrically tunable lens , 2016, SPIE Photonics Europe.

[4]  Florian Lemke,et al.  Topological in-plane polarized piezo actuation for compact adaptive lenses with aspherical correction , 2016 .

[5]  Alberto Diaspro,et al.  Enhanced volumetric imaging in 2‐photon microscopy via acoustic lens beam shaping , 2018, Journal of biophotonics.

[6]  Yeung Yam,et al.  High-speed 3D imaging based on structured illumination and electrically tunable lens , 2017 .

[7]  Xiaodong Tao,et al.  Adaptive optics confocal microscopy using direct wavefront sensing. , 2011, Optics letters.

[8]  Victoria A Griffiths,et al.  Dynamic wavefront shaping with an acousto-optic lens for laser scanning microscopy. , 2016, Optics express.

[9]  Kristen C. Maitland,et al.  Volumetric structured illumination microscopy enabled by a tunable-focus lens. , 2015, Optics letters.

[10]  Benjamin Schmid,et al.  Rapid 3D light-sheet microscopy with a tunable lens. , 2013, Optics express.

[11]  Ireneusz Grulkowski,et al.  Rapid acousto-optic focus tuning for improvement of imaging performance in confocal microscopy [Invited]. , 2018, Applied optics.

[12]  Benjamin F. Grewe,et al.  Fast two-layer two-photon imaging of neuronal cell populations using an electrically tunable lens , 2011, Biomedical optics express.

[13]  Yuan Luo,et al.  In vivo volumetric fluorescence sectioning microscopy with mechanical-scan-free hybrid illumination imaging. , 2016, Biomedical optics express.

[14]  Thomas G. Bifano,et al.  Extended depth-of-field microscopy with a high-speed deformable mirror , 2017, BiOS.

[15]  Alberto Diaspro,et al.  Simultaneous multiplane confocal microscopy using acoustic tunable lenses. , 2014, Optics express.

[16]  Ryuichi Tanimoto,et al.  High-speed microscopy with an electrically tunable lens to image the dynamics of in vivo molecular complexes. , 2015, The Review of scientific instruments.

[17]  Alberto Diaspro,et al.  Fast Inertia-Free Volumetric Light-Sheet Microscope , 2017 .

[18]  M. Hofmann,et al.  Axial scanning in confocal microscopy employing adaptive lenses (CAL). , 2014, Optics express.

[19]  Yi-Pai Huang,et al.  Fast Axial-Scanning Widefield Microscopy With Constant Magnification and Resolution , 2015, Journal of Display Technology.

[20]  Wilson,et al.  New modal wave-front sensor: a theoretical analysis , 2000, Journal of the Optical Society of America. A, Optics, image science, and vision.

[21]  T. Wilson,et al.  Adaptive aberration correction in a confocal microscope , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Na Ji,et al.  Multiplexed aberration measurement for deep tissue imaging in vivo , 2014, Nature Methods.

[23]  Ulrike Wallrabe,et al.  Volumetric HiLo microscopy employing an electrically tunable lens. , 2016, Optics express.

[24]  Martin J. Booth,et al.  Adaptive optical microscopy: the ongoing quest for a perfect image , 2014, Light: Science & Applications.

[25]  A. Mosk,et al.  Focusing coherent light through opaque strongly scattering media. , 2007, Optics letters.

[26]  Eric Betzig,et al.  Adaptive optics via pupil segmentation for high-resolution imaging in biological tissues , 2010, Nature Methods.

[27]  G. Lerosey,et al.  Controlling waves in space and time for imaging and focusing in complex media , 2012, Nature Photonics.