Optical clearing in cardiac imaging: A comparative study
Year: 2022
Authors: Olianti C., Giardini F., Lazzeri E., Costantini I., Silvestri L., Coppini R., Cerbai E., Pavone F.S., Sacconi L.
Autors Affiliation: European Lab Nonlinear Spect, I-50019 Sesto Fiorentino, Italy; CNR, Natl Inst Opt, I-50125 Florence, Italy; Univ Florence, Dept Biol, I-50019 Sesto Fiorentino, Italy; Univ Florence, Dept Phys & Astron, I-50019 Sesto Fiorentino, Italy; Univ Florence, Dept Neurosci Psychol Drugs & Child Hlth, Florence, Italy; Univ Freiburg, Fac Med, Inst Expt Cardiovasc Med, Freiburg, Germany.
Abstract: The optical clearing of the cardiac tissue has always been a challenging goal to obtain successful threedimensional reconstructions of entire hearts. Typically, the developed protocols are targeted at the clearing of the brain; cardiac tissue requires proper arrangements to the original protocols, which are usually tough and time-consuming to figure out. Here, we present the application of three different clearing methodologies on mouse hearts: uDISCO, CLARITY, and SHIELD. For each approach, we describe the required optimizations that we have developed to improve the outcome; in particular, we focus on comparing the features of the tissue after the application of each methodology, especially in terms of tissue preservation, transparency, and staining. We found that the uDISCO protocol induces strong fiber delamination of the cardiac tissue, thus reducing the reliability of structural analyses. The CLARITY protocol confers a high level of transparency to the heart and allows deep penetration of the fluorescent dyes; however, it requires long times for the clearing and the tissue loses its robustness. The SHIELD methodology, indeed, is very promising for tissue maintenance since it preserves its consistency and provides ideal transparency, but further approaches are needed to obtain homogeneous staining of the whole heart. Since the CLARITY procedure, despite the disadvantages in terms of tissue preservation and timings, is actually the most suitable approach to image labeled samples in depth, we optimized and performed the methodology also on human cardiac tissue from control hearts and hearts with hypertrophic cardiomyopathy. (c) 2021 Elsevier Ltd. All rights reserved.
Journal/Review: PROGRESS IN BIOPHYSICS & MOLECULAR BIOLOGY
Volume: 168 Pages from: 10 to: 17
More Information: This project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 95216.KeyWords: Fluorescence microscopy; Tissue clearing; Cardiovascular research; Light-sheet microscopyDOI: 10.1016/j.pbiomolbio.2021.07.012Citations: 10data from “WEB OF SCIENCE” (of Thomson Reuters) are update at: 2024-12-15References taken from IsiWeb of Knowledge: (subscribers only)