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

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.

[1]  Dimitri Perrin,et al.  Whole-Body Imaging with Single-Cell Resolution by Tissue Decolorization , 2014, Cell.

[2]  Jiří Janáček,et al.  Comparison of different tissue clearing methods and 3D imaging techniques for visualization of GFP-expressing mouse embryos and embryonic hearts , 2016, Histochemistry and Cell Biology.

[3]  W. Guido,et al.  ClearT: a detergent- and solvent-free clearing method for neuronal and non-neuronal tissue , 2013, Development.

[4]  Jyrki Lötjönen,et al.  A Comprehensive Panel of Three-Dimensional Models for Studies of Prostate Cancer Growth, Invasion and Drug Responses , 2010, PloS one.

[5]  K. Longmuir,et al.  Use of labeled tomato lectin for imaging vasculature structures , 2014, Histochemistry and Cell Biology.

[6]  J J Larsen,et al.  Femoxetine in the prophylaxis of migraine ‐ A randomised comparison with placebo , 1981, Acta neurologica Scandinavica.

[7]  Vasilis Ntziachristos,et al.  Shedding light onto live molecular targets , 2003, Nature Medicine.

[8]  Jan Huisken,et al.  Selective plane illumination microscopy techniques in developmental biology , 2009, Development.

[9]  Atsushi Miyawaki,et al.  ScaleS: an optical clearing palette for biological imaging , 2015, Nature Neuroscience.

[10]  Paul W. Frankland,et al.  Optimization of CLARITY for Clearing Whole-Brain and Other Intact Organs1,2,3 , 2015, eNeuro.

[11]  Dimitri Perrin,et al.  Advanced CUBIC protocols for whole-brain and whole-body clearing and imaging , 2015, Nature Protocols.

[12]  Yu Jin Jang,et al.  ACT-PRESTO: Rapid and consistent tissue clearing and labeling method for 3-dimensional (3D) imaging , 2016, Scientific Reports.

[13]  Ali Ertürk,et al.  Imaging Cleared Intact Biological Systems at a Cellular Level by 3DISCO , 2014, Journal of visualized experiments : JoVE.

[14]  Michael Platten,et al.  Correlated magnetic resonance imaging and ultramicroscopy (MR-UM) is a tool kit to assess the dynamics of glioma angiogenesis , 2016, eLife.

[15]  Ernst H. K. Stelzer,et al.  High-resolution deep imaging of live cellular spheroids with light-sheet-based fluorescence microscopy , 2013, Cell and Tissue Research.

[16]  Jeff W. Lichtman,et al.  Clarifying Tissue Clearing , 2015, Cell.

[17]  Anna Letizia Allegra Mascaro,et al.  A versatile clearing agent for multi-modal brain imaging , 2015, Scientific Reports.

[18]  Review: Fluorescent protein-based tumor models. , 2012, In vivo.

[19]  D H Burns,et al.  Orthogonal‐plane fluorescence optical sectioning: Three‐dimensional imaging of macroscopic biological specimens , 1993, Journal of microscopy.

[20]  Takehiro Tawara,et al.  Resource Whole-Brain Imaging with Single-Cell Resolution Using Chemical Cocktails and Computational Analysis , 2014 .

[21]  K. Deisseroth,et al.  Advanced CLARITY for rapid and high-resolution imaging of intact tissues , 2014, Nature Protocols.

[22]  Frank W. Wise,et al.  In vivo three-photon microscopy of subcortical structures within an intact mouse brain , 2012, CLEO 2012.

[23]  Frank Bradke,et al.  Three-dimensional imaging of the unsectioned adult spinal cord to assess axon regeneration and glial responses after injury , 2011, Nature Medicine.

[24]  M. Bibby,et al.  Orthotopic models of cancer for preclinical drug evaluation: advantages and disadvantages. , 2004, European journal of cancer.

[25]  N. Renier,et al.  iDISCO: A Simple, Rapid Method to Immunolabel Large Tissue Samples for Volume Imaging , 2014, Cell.

[26]  K. Alitalo,et al.  A novel multistep mechanism for initial lymphangiogenesis in mouse embryos based on ultramicroscopy , 2013, The EMBO journal.

[27]  Robert M. Hoffman,et al.  The multiple uses of fluorescent proteins to visualize cancer in vivo , 2005, Nature Reviews Cancer.

[28]  Meng-Tsen Ke,et al.  Super-Resolution Mapping of Neuronal Circuitry With an Index-Optimized Clearing Agent. , 2016, Cell reports.

[29]  R. Braren,et al.  Tumor Uptake of Anti-CD20 Fabs Depends on Tumor Perfusion , 2016, The Journal of Nuclear Medicine.

[30]  K. Deisseroth,et al.  CLARITY for mapping the nervous system , 2013, Nature Methods.

[31]  Atsushi Miyawaki,et al.  Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain , 2011, Nature Neuroscience.

[32]  Rajan P Kulkarni,et al.  Single-Cell Phenotyping within Transparent Intact Tissue through Whole-Body Clearing , 2014, Cell.

[33]  Kwanghun Chung,et al.  Simple, Scalable Proteomic Imaging for High-Dimensional Profiling of Intact Systems , 2015, Cell.

[34]  R. Muñoz-Chápuli.  Getting more CLARITY to gain the third dimension , 2013 .

[35]  Ernst H. K. Stelzer,et al.  Light sheet-based fluorescence microscopy (LSFM) for the quantitative imaging of cells and tissues , 2015, Cell and Tissue Research.

[36]  Seulki Lee,et al.  Activatable molecular probes for cancer imaging. , 2010, Current topics in medicinal chemistry.

[37]  A. Schierloh,et al.  Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain , 2007, Nature Methods.

[38]  Randolf Menzel,et al.  Three-Dimensional Reconstruction and Segmentation of Intact Drosophila by Ultramicroscopy , 2009, Front. Syst. Neurosci..

[39]  Aaron S. Andalman,et al.  Structural and molecular interrogation of intact biological systems , 2013, Nature.

[40]  P. Sharma,et al.  The future of immune checkpoint therapy , 2015, Science.

[41]  Frank Bradke,et al.  High-resolution imaging of entire organs by 3-dimensional imaging of solvent cleared organs (3DISCO) , 2013, Experimental Neurology.

[42]  S. Qadri,et al.  Oxyradical-induced GFP damage and loss of fluorescence. , 2008, International journal of biological macromolecules.

[43]  Hans-Ulrich Dodt,et al.  Formalin-Induced Fluorescence Reveals Cell Shape and Morphology in Biological Tissue Samples , 2010, PloS one.

[44]  Hiroki R Ueda,et al.  Whole-body and Whole-Organ Clearing and Imaging Techniques with Single-Cell Resolution: Toward Organism-Level Systems Biology in Mammals. , 2016, Cell chemical biology.

[45]  Vasilis Ntziachristos,et al.  Multispectral fluorescence ultramicroscopy: three-dimensional visualization and automatic quantification of tumor morphology, drug penetration, and antiangiogenic treatment response. , 2014, Neoplasia.

[46]  W. Kalender,et al.  Dynamic Contrast-Enhanced Micro–Computed Tomography Correlates With 3-Dimensional Fluorescence Ultramicroscopy in Antiangiogenic Therapy of Breast Cancer Xenografts , 2014, Investigative radiology.

[47]  Zhen Cheng,et al.  Near-infrared fluorescent nanoprobes for cancer molecular imaging: status and challenges. , 2010, Trends in molecular medicine.

[48]  F. Del Bene,et al.  Optical Sectioning Deep Inside Live Embryos by Selective Plane Illumination Microscopy , 2004, Science.

[49]  R. Weiler,et al.  Chemical Clearing and Dehydration of GFP Expressing Mouse Brains , 2012, PloS one.

[50]  P. Santi,et al.  Light Sheet Fluorescence Microscopy , 2011, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[51]  Hiroki R Ueda,et al.  Chemical Principles in Tissue Clearing and Staining Protocols for Whole-Body Cell Profiling. , 2016, Annual review of cell and developmental biology.

[52]  David A. Tuveson,et al.  Maximizing mouse cancer models , 2007, Nature Reviews Cancer.

[53]  Axel Hoos,et al.  Non-alcoholic steatohepatitis: emerging molecular targets and therapeutic strategies , 2016, Nature Reviews Drug Discovery.

[54]  Hans-Ulrich Dodt,et al.  Resolution of Ultramicroscopy and Field of View Analysis , 2009, PloS one.

[55]  J Wu,et al.  Intrinsic fluorescence spectroscopy in turbid media: disentangling effects of scattering and absorption. , 2001, Applied optics.

[56]  E. Susaki,et al.  Whole-Brain Imaging with Single-Cell Resolution Using Chemical Cocktails and Computational Analysis , 2014, Cell.

[57]  Philipp J. Keller,et al.  Shedding light on the system: studying embryonic development with light sheet microscopy. , 2011, Current Opinion in Genetics and Development.

[58]  J. Wolchok,et al.  Immune modulation in cancer with antibodies. , 2014, Annual review of medicine.

[59]  Hisataka Kobayashi,et al.  Target-cancer-cell-specific activatable fluorescence imaging probes: rational design and in vivo applications. , 2011, Accounts of chemical research.

[60]  Hans-Ulrich Dodt,et al.  Ultramicroscopy: light-sheet-based microscopy for imaging centimeter-sized objects with micrometer resolution. , 2013, Cold Spring Harbor protocols.

[61]  Takeshi Imai,et al.  SeeDB: a simple and morphology-preserving optical clearing agent for neuronal circuit reconstruction , 2013, Nature Neuroscience.

[62]  Werner Spalteholz,et al.  Über das Durchsichtigmachen von menschlichen und tierischen Präparaten : nebst Anhang: Über Knochenfärbung , 2022 .

[63]  H. Dodt,et al.  3D-reconstruction of blood vessels by ultramicroscopy , 2009, Organogenesis.

[64]  Charless C. Fowlkes,et al.  Whole-body tissue stabilization and selective extractions via tissue-hydrogel hybrids for high-resolution intact circuit mapping and phenotyping , 2015, Nature Protocols.

[65]  Frank Bradke,et al.  Three-dimensional imaging of solvent-cleared organs using 3DISCO , 2012, Nature Protocols.

[66]  Bernard Choi,et al.  Optical properties of mouse brain tissue after optical clearing with FocusClear™ , 2015, Journal of biomedical optics.

[67]  Bernard Choi,et al.  High-resolution visualization of mouse cardiac microvasculature using optical histology. , 2013, Biomedical optics express.

[68]  H. Siedentopf,et al.  Uber Sichtbarmachung und Größenbestimmung ultramikoskopischer Teilchen, mit besonderer Anwendung auf Goldrubingläser , 1902 .

[69]  Tianzi Jiang,et al.  Scalable and DiI-compatible optical clearance of the mammalian brain , 2015, Front. Neuroanat..

[70]  Vasilis Ntziachristos,et al.  Looking and listening to light: the evolution of whole-body photonic imaging , 2005, Nature Biotechnology.

[71]  Frank Bradke,et al.  A simple method for 3D analysis of immunolabeled axonal tracts in a transparent nervous system. , 2014, Cell reports.

[72]  G. Kalman.  Nonlinear oscillations and nonstationary flow in a zero temperature plasma: Part I. Initial and boundary value problems , 1960 .

[73]  Martin K. Schwarz,et al.  Fluorescent-Protein Stabilization and High-Resolution Imaging of Cleared, Intact Mouse Brains , 2015, PloS one.

[74]  I. Tannock,et al.  Drug penetration in solid tumours , 2006, Nature Reviews Cancer.

[75]  Meng-Tsen Ke,et al.  Optical Clearing of Fixed Brain Samples Using SeeDB , 2014, Current protocols in neuroscience.

[76]  F. Jöbsis,et al.  Non-invasive, infra-red monitoring of cerebral O2 sufficiency, bloodvolume, HbO2-Hb shifts and bloodflow. , 1977, Acta neurologica Scandinavica. Supplementum.

[77]  H. Dodt,et al.  Ultramicroscopy: 3D reconstruction of large microscopical specimens , 2008, Journal of biophotonics.

[78]  Kwanghun Chung,et al.  Light microscopy mapping of connections in the intact brain , 2013, Trends in Cognitive Sciences.

[79]  Farooq Azam,et al.  Thin laser light sheet microscope for microbial oceanography. , 2002, Optics express.

[80]  R. Hoffman.  Application of GFP imaging in cancer , 2014, Laboratory Investigation.

[81]  V. Ntziachristos.  Fluorescence molecular imaging. , 2006, Annual review of biomedical engineering.