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Microfabricated devices for single objective single plane illumination microscopy (SoSPIM)

Elisa Zagato (UGent) , Toon Brans (UGent) , Steven Verstuyft (UGent) , Dries Van Thourhout (UGent) , Jeroen Missinne (UGent) , Geert Van Steenberge (UGent) , Jo Demeester (UGent) , Stefaan De Smedt (UGent) , Katrien Remaut (UGent) , Kristiaan Neyts (UGent) , et al.
(2017) OPTICS EXPRESS. 25(3). p.1732-1745
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Abstract
Light sheet microscopy is a relatively new form of fluorescence microscopy that has been receiving a lot of attention recently. The strong points of the technique, such as high signal to noise ratio and its reduced photodamage of fluorescently labelled samples, come from its unique feature to illuminate only a thin plane in the sample that coincides with the focal plane of the detection lens. Typically this requires two closely positioned perpendicular objective lenses, one for detection and one for illumination. Apart from the fact that this special configuration of objective lenses is incompatible with standard microscope bodies, it is particularly problematic for high-resolution lenses which typically have a short working distance. To address these issues we developed sample holders with an integrated micromirror to perform single lens light sheet microscopy, also known as single objective single plane illumination microscopy (SoSPIM). The first design is based on a wet-etched silicon substrate, the second on a microfabricated polished polymer plug. We achieved an on-chip light sheet thickness of 2.3 mu m (FWHM) at 638 nm with the polymer micromirror and of 1.7 mu m (FWHM) at 638 nm with the silicon micromirror, comparable to reported light sheet thicknesses obtained on dedicated light sheet microscopes. A marked contrast improvement was obtained with both sample holders as compared to classic epi-fluorescence microscopy. In order to evaluate whether this technology could be made available on a larger scale, in a next step we evaluated the optical quality of inexpensive replicas from both types of master molds. We found that replicas from the polished polymer based mold have an optical quality close to that of the master component, while replicas from the silicon based mold were of slightly lower but still acceptable quality. The suitability of the replicated polymer based sample holder for single-lens light sheet microscopy was finally demonstrated by imaging breast cancer spheroids. (C) 2017 Optical Society of America
Keywords
LIGHT-SHEET MICROSCOPY, PARTICLE TRACKING, EXCITATION, HEART

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MLA
Zagato, Elisa, et al. “Microfabricated Devices for Single Objective Single Plane Illumination Microscopy (SoSPIM).” OPTICS EXPRESS, vol. 25, no. 3, 2017, pp. 1732–45, doi:10.1364/oe.25.001732.
APA
Zagato, E., Brans, T., Verstuyft, S., Van Thourhout, D., Missinne, J., Van Steenberge, G., … Braeckmans, K. (2017). Microfabricated devices for single objective single plane illumination microscopy (SoSPIM). OPTICS EXPRESS, 25(3), 1732–1745. https://doi.org/10.1364/oe.25.001732
Chicago author-date
Zagato, Elisa, Toon Brans, Steven Verstuyft, Dries Van Thourhout, Jeroen Missinne, Geert Van Steenberge, Jo Demeester, et al. 2017. “Microfabricated Devices for Single Objective Single Plane Illumination Microscopy (SoSPIM).” OPTICS EXPRESS 25 (3): 1732–45. https://doi.org/10.1364/oe.25.001732.
Chicago author-date (all authors)
Zagato, Elisa, Toon Brans, Steven Verstuyft, Dries Van Thourhout, Jeroen Missinne, Geert Van Steenberge, Jo Demeester, Stefaan De Smedt, Katrien Remaut, Kristiaan Neyts, and Kevin Braeckmans. 2017. “Microfabricated Devices for Single Objective Single Plane Illumination Microscopy (SoSPIM).” OPTICS EXPRESS 25 (3): 1732–1745. doi:10.1364/oe.25.001732.
Vancouver
1.
Zagato E, Brans T, Verstuyft S, Van Thourhout D, Missinne J, Van Steenberge G, et al. Microfabricated devices for single objective single plane illumination microscopy (SoSPIM). OPTICS EXPRESS. 2017;25(3):1732–45.
IEEE
[1]
E. Zagato et al., “Microfabricated devices for single objective single plane illumination microscopy (SoSPIM),” OPTICS EXPRESS, vol. 25, no. 3, pp. 1732–1745, 2017.
@article{8519654,
  abstract     = {{Light sheet microscopy is a relatively new form of fluorescence microscopy that has been receiving a lot of attention recently. The strong points of the technique, such as high signal to noise ratio and its reduced photodamage of fluorescently labelled samples, come from its unique feature to illuminate only a thin plane in the sample that coincides with the focal plane of the detection lens. Typically this requires two closely positioned perpendicular objective lenses, one for detection and one for illumination. Apart from the fact that this special configuration of objective lenses is incompatible with standard microscope bodies, it is particularly problematic for high-resolution lenses which typically have a short working distance. To address these issues we developed sample holders with an integrated micromirror to perform single lens light sheet microscopy, also known as single objective single plane illumination microscopy (SoSPIM). The first design is based on a wet-etched silicon substrate, the second on a microfabricated polished polymer plug. We achieved an on-chip light sheet thickness of 2.3 mu m (FWHM) at 638 nm with the polymer micromirror and of 1.7 mu m (FWHM) at 638 nm with the silicon micromirror, comparable to reported light sheet thicknesses obtained on dedicated light sheet microscopes. A marked contrast improvement was obtained with both sample holders as compared to classic epi-fluorescence microscopy. In order to evaluate whether this technology could be made available on a larger scale, in a next step we evaluated the optical quality of inexpensive replicas from both types of master molds. We found that replicas from the polished polymer based mold have an optical quality close to that of the master component, while replicas from the silicon based mold were of slightly lower but still acceptable quality. The suitability of the replicated polymer based sample holder for single-lens light sheet microscopy was finally demonstrated by imaging breast cancer spheroids. (C) 2017 Optical Society of America}},
  author       = {{Zagato, Elisa and Brans, Toon and Verstuyft, Steven and Van Thourhout, Dries and Missinne, Jeroen and Van Steenberge, Geert and Demeester, Jo and De Smedt, Stefaan and Remaut, Katrien and Neyts, Kristiaan and Braeckmans, Kevin}},
  issn         = {{1094-4087}},
  journal      = {{OPTICS EXPRESS}},
  keywords     = {{LIGHT-SHEET MICROSCOPY,PARTICLE TRACKING,EXCITATION,HEART}},
  language     = {{eng}},
  number       = {{3}},
  pages        = {{1732--1745}},
  title        = {{Microfabricated devices for single objective single plane illumination microscopy (SoSPIM)}},
  url          = {{http://dx.doi.org/10.1364/oe.25.001732}},
  volume       = {{25}},
  year         = {{2017}},
}

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