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Biomed Opt Express. 2017 Aug 1;8(9):3918-3937. doi: 10.1364/BOE.8.003918. eCollection 2017 Sep 1.

sideSPIM - selective plane illumination based on a conventional inverted microscope.

Author information

1
Laboratory for Fluorescence Dynamics, Department of Biomedical Engineering, University of California Irvine, Irvine, CA, USA.
2
phedde@uci.edu.
3
Área de Investigación Respiratoria, Departamento de Fisiopatología, Hospital de Clínicas, Facultad de Medicina, Universidad de la República, Uruguay.
4
Analytical Biochemistry and Proteomics Unit, Institut Pasteur of Montevideo, Montevideo, Uruguay.
5
Department of Physics and Astronomy, University of California Irvine, Irvine, CA, USA.
6
Department of Molecular Biology and Biochemistry, University of California Irvine, Irvine, CA, USA.

Abstract

Previously described selective plane illumination microscopy techniques typically offset ease of use and sample handling for maximum imaging performance or vice versa. Also, to reduce cost and complexity while maximizing flexibility, it is highly desirable to implement light sheet microscopy such that it can be added to a standard research microscope instead of setting up a dedicated system. We devised a new approach termed sideSPIM that provides uncompromised imaging performance and easy sample handling while, at the same time, offering new applications of plane illumination towards fluidics and high throughput 3D imaging of multiple specimen. Based on an inverted epifluorescence microscope, all of the previous functionality is maintained and modifications to the existing system are kept to a minimum. At the same time, our implementation is able to take full advantage of the speed of the employed sCMOS camera and piezo stage to record data at rates of up to 5 stacks/s. Additionally, sample handling is compatible with established methods and switching magnification to change the field of view from single cells to whole organisms does not require labor intensive adjustments of the system.

KEYWORDS:

(110.6880) Three-dimensional image acquisition; (170.2520) Fluorescence microscopy; (180.6900) Three-dimensional microscopy; (220.1000) Aberration compensation; (220.2945) Illumination design

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