Multiview deconvolution approximation multiphoton microscopy of tissues and zebrafish larvae

Scientific Reports, vol. 11, pp. 10160

Abstract

Imaging in three dimensions is necessary for thick tissues and small organisms. This is possible with tomographic optical microscopy techniques such as confocal, multiphoton and light sheet microscopy. All these techniques suffer from anisotropic resolution and limited penetration depth. In the past, Multiview microscopy-imaging the sample from different angles followed by 3D image reconstruction-was developed to address this issue for light sheet microscopy based on fluorescence signal. In this study we applied this methodology to accomplish Multiview imaging with multiphoton microscopy based on fluorescence and additionally second harmonic signal from myosin and collagen. It was shown that isotropic resolution was achieved, the entirety of the sample was visualized, and interference artifacts were suppressed allowing clear visualization of collagen fibrils and myofibrils. This method can be applied to any scanning microscopy technique without microscope modifications. It can be used for imaging tissue and whole mount small organisms such as heart tissue, and zebrafish larva in 3D, label-free or stained, with at least threefold axial resolution improvement which can be significant for the accurate quantification of small 3D structures.

Authors 8

1. Dimitrios Kapsokalyvas corresponding Aachen Institute for Molecular Cardiovascular Research

   Maastricht University · Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Department of Genetics and Cell Biology, Faculty of Health, Medicine and Life Sciences (FHML), Maastricht University, Maastricht, the Netherlands. d.kapsokalyvas@maastrichtuniversity.nl

   Institute for Molecular Cardiovascular Research (IMCAR), University Hospital RWTH Aachen University, Aachen, Germany. d.kapsokalyvas@maastrichtuniversity.nl

   Department of Genetics and Cell Biology, Faculty of Health, Medicine and Life Sciences (FHML), Maastricht University, Maastricht, the Netherlands

2. Rodrigo A. Rosas

   Maastricht University

   Affiliation as printed

   Department of Genetics and Cell Biology, Faculty of Health, Medicine and Life Sciences (FHML), Maastricht University, Maastricht, the Netherlands

3. Rob W. A. Janssen

   Maastricht University

   Affiliation as printed

   Department of Genetics and Cell Biology, Faculty of Health, Medicine and Life Sciences (FHML), Maastricht University, Maastricht, the Netherlands

4. Jo M. Vanoevelen

   Maastricht University

   Affiliation as printed

   Department of Genetics and Cell Biology, Faculty of Health, Medicine and Life Sciences (FHML), Maastricht University, Maastricht, the Netherlands

5. Miranda Nabben

   Maastricht University

   Affiliation as printed

   Department of Genetics and Cell Biology, Faculty of Health, Medicine and Life Sciences (FHML), Maastricht University, Maastricht, the Netherlands

6. Martin Strauch Aachen Institute of Imaging & Computer Vision

   RWTH Aachen University

   Affiliation as printed

   Institute of Imaging and Computer Vision, RWTH Aachen University, Aachen, Germany

7. Dorit Merhof Aachen Institute of Imaging & Computer Vision

   RWTH Aachen University

   Affiliation as printed

   Institute of Imaging and Computer Vision, RWTH Aachen University, Aachen, Germany

8. Marc A. M. J. van Zandvoort Aachen Institute for Molecular Cardiovascular Research

   Maastricht University · Universitätsklinikum Aachen · RWTH Aachen University

   Affiliation as printed

   Department of Genetics and Cell Biology, Faculty of Health, Medicine and Life Sciences (FHML), Maastricht University, Maastricht, the Netherlands

   Institute for Molecular Cardiovascular Research (IMCAR), University Hospital RWTH Aachen University, Aachen, Germany