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Hepatic Vitamin A Content Investigation Using Coherent Anti‐Stokes Raman Scattering Microscopy |
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| Authors: | Fisseha Bekele Legesse Dr. Sandro Heuke Dr. Kerstin Galler Patrick Hoffmann Prof. Dr. Michael Schmitt Prof. Dr. Ute Neugebauer Prof. Dr. Michael Bauer Prof. Dr. Jürgen Popp |
| Affiliation: | 1. Institute of Physical Chemistry and Abbe Center of Photonics, Friedrich Schiller University Jena, Jena, Germany;2. Leibniz Institute of Photonic Technology (IPHT) Jena e.V., Jena, Germany;3. Center for Sepsis Control and Care, Jena University Hospital, Jena, Germany;4. Department of Anesthesiology and Intensive Care Medicine, Jena University Hospital, Jena, Germany |
| Abstract: | Standard techniques for examining the distribution of vitamin A in liver either require staining or lead to rapid photobleaching of the molecule. A potentially better alternative approach is to use coherent anti‐Stokes Raman scattering (CARS) microscopy; a fast, label‐free, non‐disruptive imaging method that provides contrast based on molecular vibrations. This contribution evaluates the viability of CARS microscopy for imaging vitamin A within thick hepatic tissue under physiological conditions by tuning into its characteristic vibrational band in the fingerprint region. Additional information about the morphology and architecture of the tissue was acquired using second harmonic generation (SHG) and multi‐photon excited fluorescence (MPEF) to help mapping the intra‐lobular positions of the vitamin A droplets. We demonstrate the capability of our multimodal imaging framework to selectively image lipid‐soluble vitamin A droplets deep in bulk liver tissue with a high contrast while co‐registering a complementary morphological background that clearly visualizes hepatic lobules. The results obtained envisage the good prospect of the technique for in vivo studies assessing vitamin A distribution heterogeneity and how it is affected by the progression of hepatic diseases. |
| Keywords: | coherent anti-Stokes Raman scattering fluorescence imaging hepatic stellate cells vibrational spectroscopy vitamin A |