We have developed an imaging technique which combines selective plane illumination microscopy with time-domain fluorescence lifetime imaging microscopy (SPIM-FLIM) for three-dimensional volumetric imaging of cleared mouse brains with micro- to mesoscopic resolution. The main features of the microscope include a wavelength-adjustable pulsed laser source (Ti:sapphire) (near-infrared) laser, a BiBO frequency-doubling photonic crystal, a liquid chamber, an electrically focus-tunable lens, a cuvette based sample holder, and an air (dry) objective lens. The performance of the system was evaluated with a lifetime reference dye and micro-bead phantom measurements. Intensity and lifetime maps of three-dimensional human embryonic kidney (HEK) cell culture samples and cleared mouse brain samples expressing green fluorescent protein (GFP) (donor only) and green and red fluorescent protein [positive Förster (fluorescence) resonance energy transfer] were acquired. The results show that the SPIM-FLIM system can be used for sample sizes ranging from single cells to whole mouse organs and can serve as a powerful tool for medical and biological research.
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May 2018
Research Article|
May 14 2018
Selective plane illumination microscopy (SPIM) with time-domain fluorescence lifetime imaging microscopy (FLIM) for volumetric measurement of cleared mouse brain samples
Tsukasa Funane
;
Tsukasa Funane
1
Department of Neurology, Harvard Medical School, Massachusetts General Hospital
, 114 16th Street, Charlestown, Massachusetts 02129, USA
2
Hitachi, Ltd., Research & Development Group, Center for Exploratory Research
, Hatoyama, Saitama 350-0395, Japan
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Steven S. Hou;
Steven S. Hou
1
Department of Neurology, Harvard Medical School, Massachusetts General Hospital
, 114 16th Street, Charlestown, Massachusetts 02129, USA
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Katarzyna Marta Zoltowska
;
Katarzyna Marta Zoltowska
1
Department of Neurology, Harvard Medical School, Massachusetts General Hospital
, 114 16th Street, Charlestown, Massachusetts 02129, USA
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Susanne J. van Veluw;
Susanne J. van Veluw
1
Department of Neurology, Harvard Medical School, Massachusetts General Hospital
, 114 16th Street, Charlestown, Massachusetts 02129, USA
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Oksana Berezovska;
Oksana Berezovska
1
Department of Neurology, Harvard Medical School, Massachusetts General Hospital
, 114 16th Street, Charlestown, Massachusetts 02129, USA
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Anand T. N. Kumar;
Anand T. N. Kumar
3
Athinoula A. Martinos Center for Biomedical Imaging, Harvard Medical School, Massachusetts General Hospital
, 149 13th Street, Charlestown, Massachusetts 02129, USA
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Brian J. Bacskai
Brian J. Bacskai
a)
1
Department of Neurology, Harvard Medical School, Massachusetts General Hospital
, 114 16th Street, Charlestown, Massachusetts 02129, USA
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a)
E-mail: [email protected]
Rev. Sci. Instrum. 89, 053705 (2018)
Article history
Received:
December 10 2017
Accepted:
April 24 2018
Citation
Tsukasa Funane, Steven S. Hou, Katarzyna Marta Zoltowska, Susanne J. van Veluw, Oksana Berezovska, Anand T. N. Kumar, Brian J. Bacskai; Selective plane illumination microscopy (SPIM) with time-domain fluorescence lifetime imaging microscopy (FLIM) for volumetric measurement of cleared mouse brain samples. Rev. Sci. Instrum. 1 May 2018; 89 (5): 053705. https://doi.org/10.1063/1.5018846
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