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Label-free 3D molecular imaging of living tissues using Raman spectral projection tomography.
Stepula, Elzbieta; Walther, Anders R; Jensen, Magnus; Mehrotra, Dev R; Yuan, Mu H; Pedersen, Simon V; Kumar, Vishal; Gentleman, Eileen; Albro, Michael B; Hedegaard, Martin A B; Bergholt, Mads S.
Afiliación
  • Stepula E; Centre for Craniofacial & Regenerative Biology, King's College London, London, UK.
  • Walther AR; SDU Chemical Engineering, Faculty of Engineering, University of Southern Denmark, Odense, Denmark.
  • Jensen M; Centre for Craniofacial & Regenerative Biology, King's College London, London, UK.
  • Mehrotra DR; Department of Mechanical Engineering, Boston University, Boston, MA, USA.
  • Yuan MH; Centre for Craniofacial & Regenerative Biology, King's College London, London, UK.
  • Pedersen SV; SDU Biotechnology, Faculty of Engineering, University of Southern Denmark, Odense, Denmark.
  • Kumar V; Centre for Craniofacial & Regenerative Biology, King's College London, London, UK.
  • Gentleman E; Centre for Craniofacial & Regenerative Biology, King's College London, London, UK.
  • Albro MB; Department of Biomedical Sciences, University of Lausanne, Lausanne, Switzerland.
  • Hedegaard MAB; Department of Mechanical Engineering, Boston University, Boston, MA, USA.
  • Bergholt MS; SDU Chemical Engineering, Faculty of Engineering, University of Southern Denmark, Odense, Denmark. marhe@igt.sdu.dk.
Nat Commun ; 15(1): 7717, 2024 Sep 09.
Article en En | MEDLINE | ID: mdl-39251593
ABSTRACT
The ability to image tissues in three dimensions (3D) with label-free molecular contrast at the mesoscale would be a valuable capability in biology and biomedicine. Here, we introduce Raman spectral projection tomography (RSPT) for volumetric molecular imaging with optical sub-millimeter spatial resolution. We have developed a RSPT imaging instrument capable of providing 3D molecular contrast in transparent and semi-transparent samples. We also created a computational pipeline for multivariate reconstruction to extract label-free spatial molecular information from Raman projection data. Using these tools, we demonstrate imaging and visualization of phantoms of various complex shapes with label-free molecular contrast. Finally, we apply RSPT as a tool for imaging of molecular gradients and extracellular matrix heterogeneities in fixed and living tissue-engineered constructs and explanted native cartilage tissues. We show that there exists a favorable balance wherein employing Raman spectroscopy, with its advantages in live cell imaging and label-free molecular contrast, outweighs the reduction in imaging resolution and blurring caused by diffuse photon propagation. Thus, RSPT imaging opens new possibilities for label-free molecular monitoring of tissues.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectrometría Raman / Fantasmas de Imagen / Imagenología Tridimensional / Imagen Molecular Límite: Animals / Humans Idioma: En Revista: Nat Commun Asunto de la revista: BIOLOGIA / CIENCIA Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Espectrometría Raman / Fantasmas de Imagen / Imagenología Tridimensional / Imagen Molecular Límite: Animals / Humans Idioma: En Revista: Nat Commun Asunto de la revista: BIOLOGIA / CIENCIA Año: 2024 Tipo del documento: Article Pais de publicación: Reino Unido