Non-linear imaging and characterization of atherosclerotic arterial tissue using combined two photon fluorescence, second-harmonic generation and CARS microscopy

Year: 2014

Authors: Cicchi R., Matthaeus C., Meyer T., Lattermann A., Dietzek B., Brehm BR., Popp J., Pavone FS.

Autors Affiliation: National Institute of Optics, National Research Council (INO-CNR), Largo E. Fermi 6 – 50125,
Florence, Italy;
European Laboratory for Non-Linear Spectroscopy (LENS), University of Florence, Via Nello Carrara, 1 – 50019, Sesto Fiorentino (Firenze), Italy;
Institute of Photonic Technology (IPHT-Jena), Albert Einstein Straße 9, 07745 Jena, Germany;
Institute of Pathology, Department of Neuropathology, Jena University Hospital – Friedrich-Schiller-University, Erlanger Allee 101, 07740 Jena, Germany;
Catholic Clinic – Koblenz, Internal Medicine & Cardiology, Rudolf Virchow Str. 9, 56073 Koblenz, Germany;
Institute of Physical Chemistry and Abbe Center of Photonics– Friedrich Schiller University Jena, Helmholtzweg 4, 07743 Jena, Germany;
Department of Physics, University of Florence, Via Giovanni Sansone 1 – 50019, Sesto Fiorentino, Italy

Abstract: Atherosclerosis is among the most widespread cardiovascular diseases and one of the leading cause of death in the Western World. Characterization of arterial tissue in atherosclerotic condition is extremely interesting from the diagnostic point of view. Routinely used diagnostic methods, such as histopathological examination, are limited to morphological analysis of the examined tissues, whereas an exhaustive characterization requires a morpho-functional approach. Multimodal non-linear microscopy has the potential to bridge this gap by providing morpho-functional information on the examined tissues in a label-free way. Here we employed multiple non-linear microscopy techniques, including CARS, TPF, and SHG to provide intrinsic optical contrast from various tissue components in both arterial wall and atherosclerotic plaques. CARS and TPF microscopy were used to respectively image lipid depositions within plaques and elastin in the arterial wall. Cholesterol deposition in the lumen and collagen in the arterial wall were selectively imaged by SHG microscopy and distinguished by forward-backward SHG ratio. Image pattern analysis allowed characterizing collagen organization in different tissue regions. Different values of fiber mean size, distribution and anisotropy are calculated for lumen and media prospectively allowing for automated classification of atherosclerotic lesions. The presented method represents a promising diagnostic tool for evaluating atherosclerotic tissue and has the potential to find a stable place in clinical setting as well as to be applied in vivo in the near future.

Conference title: Photonics West – BIOS 2014
Place: San Francisco, US

More Information: The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007- 2013) under grant agreements number 228334 and 284464, and from the Italian Ministry for Education, University and Research in the framework of the Flagship Project NANOMAX. Furthermore, financial support by Ente Cassa di Risparmio di Firenze ( private foundation) is acknowledged. We are grateful for support by the European network of excellence Photonics4Life (P4L).
KeyWords: Characterization; Coherent scattering; Collagen; Deposition; Diseases; Histology, Atherosclerotic lesions; Automated classification; Cardio-vascular disease; Histopathological examinations; Intrinsic optical contrasts; Nonlinear microscopy; Scanning microscopy; Tissue characterization, Tissue
DOI: 10.1117/12.2037018

Citations: 2
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