Abstract
Point-scanning-based microscopy systems require combination of axial and lateral scanning to obtain three-dimensional (3D) data. Axial scanning was commonly achieved by mechanical displacement of the objective or the sample. To improve, various adaptive lens-based solutions have been reported to circumvent the need for mechanically moving parts. The lateral scanning is predominantly implemented using galvanometric mirrors. Although the performance of such devices is flawless, they require bulky, folded beam-paths that make their incorporation in compact hand-held devices challenging. Recently, we introduced an adaptive prism as a transmissive device that enables lateral scanning. We demonstrate the first all-adaptive 3D scanning in laser scanning microscopes employing a compact in-line transmission geometry without mechanically moving parts and beam folding, combining an adaptive lens and a novel adaptive prism. Characterization of the all-adaptive microscope performance shows a lateral tuning range of approximately X = Y = 130 μm and an axial tuning range of about Z = 500 μm. We successfully demonstrate 3D raster scanning of the fluorescence of a thyroid of a zebrafish embryo.
Highlights
Adaptive optics technology has been employed in various systems to improve imaging properties,[1] correct aberrations,[2] and perform self-calibration.[3]
More sophisticated adaptive lenses with 2 degrees of freedom enabled simultaneously tuning the focal position and correcting the spherical aberrations. This lens is committed to correct for specimen-induced spherical aberrations that occur in deep tissue applications[2] and systematic scan-induced aberrations.[12]
The adaptive lens used in our all-adaptive scanning microscope consists of a transparent polydimethylsiloxan membrane, which is embedded in an annular piezo-bending actuator
Summary
Adaptive optics technology has been employed in various systems to improve imaging properties,[1] correct aberrations,[2] and perform self-calibration.[3]. Several approaches employing adaptive lenses[11,12] have been introduced to realize axial scanning without the need for any mechanically moving parts in a variety of microscopes, such as confocal microscopy,[2,13] two-photon microscopy,[14,15] light-sheet microscopy,[16,17,18] structured illumination microscopy,[19,20] and standard wide-field microscopy.[21,22] More sophisticated adaptive lenses with 2 degrees of freedom enabled simultaneously tuning the focal position and correcting the spherical aberrations. The lateral scan is accomplished by mechanical translation of the sample or by reflective galvanometric mirrors.[23] The direction of the reflected beam can be regulated by turning the mirror This type of scanner has a high resolution, good repeatability, and low drift values. To demonstrate the ability for biomedical 3D imaging of our approach, we conduct raster scanning of the thyroid of a cultured zebrafish embryo
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
