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Deep tissue imaging: a review from a preclinical cancer research perspective

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Abstract

This review delves into the rapidly evolving field of deep tissue imaging at cellular resolution, reviewing popular tissue clearing and staining methods in combination with light-sheet fluorescence microscopy (LSFM) including quantification and three-dimensional visualization tools, the field of applications and perspective, particularly with the focus on preclinical cancer research and drug development. The LSFM technique presented here allows an extremely fast optical sectioning for three-dimensional reconstruction of centimeter-sized tissue samples at cellular resolution. However, optical clearing methods are required to receive optical transparent tissue. Application of either tissue autofluorescence, in vivo fluorescence labeling, endogenous fluorescence or ex vivo whole-mount immunolabeling enables three-dimensional in situ visualization of morphological and functional features of unsectioned whole-mount tissue samples. This powerful and innovative imaging technique opens up a new dimension of tissue analysis providing detailed and comprehensive insights into biology. It enables the investigation of normal and pathological tissue features and disease progression and allows precise monitoring of potential therapeutic interventions in intact biological tissue on a cellular level.

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Acknowledgments

The authors gratefully acknowledge the financial support of the Deutsche Forschungsgemeinschaft (SFB 824 TP Z02). The authors AF and AW would like to thank Ulrike Buchholz, Claudia-Mareike Pflüger and Andreas Voss for excellent technical assistance. The author MD would like to thank Hadassah Sade, Thomas Pöschinger and Frank Herting for helpful discussion and constant support and the Roche Postdoc Fellowship Program for financing this work.

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Authors and Affiliations

  1. Research Unit Analytical Pathology, Helmholtz Zentrum München, Neuherberg, Germany

    Annette Feuchtinger & Axel Walch

  2. Institute of Pathology, Technische Universität München, Munich, Germany

    Annette Feuchtinger

  3. Discovery Oncology, Pharma Research and Early Development (pRED), Roche Diagnostics GmbH, Penzberg, Germany

    Michael Dobosz

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  1. Annette Feuchtinger
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  2. Axel Walch
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Correspondence to Axel Walch.

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The authors AF and AW declare no conflict of interest. The author MD is an employee of Roche Diagnostics GmbH.

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This review article is dedicated to Professor emeritus Detlev Drenckhahn for his outstanding achievements in the field of histochemistry and cell biology.

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Morphology of a mouse lung (cleared according to 3DISCO) based on autofluorescence signals visualized by arivis Vision4D (AVI 140067 kb)

Video 2

2D visualization of virtual tissue slices obtained from different fluorescence channels. First channel: Tumor morphology (autofluorescence, gray). Second channel: Tumor vasculature (Lectin-Alexa647, green). Third channel: Antibody penetration (Trastuzumab-Alexa750, red) (MP4 26017 kb)

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Feuchtinger, A., Walch, A. & Dobosz, M. Deep tissue imaging: a review from a preclinical cancer research perspective. Histochem Cell Biol 146, 781–806 (2016). https://doi.org/10.1007/s00418-016-1495-7

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