Attenuation artifacts in light sheet fluorescence microscopy corrected by OPTiSPIM

Jürgen Mayer; Alexandre Robert-Moreno; James Sharpe; Jim Swoger

doi:10.1038/s41377-018-0068-z

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Attenuation artifacts in light sheet fluorescence microscopy corrected by OPTiSPIM

Original Articles

- Attenuation artifacts in light sheet fluorescence microscopy corrected by OPTiSPIM
- Light: Science & Applications Vol. 7, Issue 6, Pages: 703-715(2018)
- Affiliations：
  
  1.Centre for Genomic Regulation (CRG), The Barcelona Institute of Science and Technology, Dr. Aiguader 88, 08003, Barcelona, Spain
  2.Universitat Pompeu Fabra (UPF), Barcelona, Spain
  3.Institució Catalana de Recerca i Estudis Avançats (ICREA), Pg. Lluis Companys 23, 08010, Barcelona, Spain
  4.Present address: European Molecular Biology Laboratory (EMBL), Dr. Aiguader 88, 08003 Barcelona, Spain
- Author bio：
  
  Jim Swoger (jim.swoger@embl.es)
- Funds：
- DOI：10.1038/s41377-018-0068-z
  CLC：
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Mayer, J. et al. Attenuation artifacts in light sheet fluorescence microscopy corrected by OPTiSPIM. Light: Science & Applications, 7, 703-715 (2018).
DOI：

Mayer, J. et al. Attenuation artifacts in light sheet fluorescence microscopy corrected by OPTiSPIM. Light: Science & Applications, 7, 703-715 (2018). DOI： 10.1038/s41377-018-0068-z.

摘要

Abstract

Light sheet fluorescence microscopy (LSFM) is rapidly becoming an essential technology for mesoscopic imaging of samples such as embryos and adult mouse organs. However, LSFM can suffer from optical artifacts for which there is no intrinsic solution. The attenuation of light due to absorbing material causes "shadow" artifacts along both the illumination and detection paths. Several approaches have been introduced to reduce this problem, including scanning illumination and multi-view imaging. However, neither of these approaches completely eliminates the problem. If the distribution of the absorbing material is complex, shadows cannot be avoided. We introduce a new approach that relies on multi-modal integration of two very different mesoscopic techniques. Unlike LSFM, optical projection tomography (OPT) can operate in transmission mode to create a voxel map of the 3D distribution of the sample's optical attenuation. Here, we demonstrate a hybrid instrument (OPTiSPIM) that can quantify this attenuation and use the information to correct the shadow artifacts of LSFM.

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references

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