Mesoscopic Optical Imaging of Whole Mouse Heart
Year: 2021
Authors: Giardini F., Lazzeri E., Olianti C., Beconi G., Costantini I., Silvestri L., Cerbai E., Pavone FS., Sacconi L.
Autors Affiliation: Univ Florence, European Lab Nonlinear Spect, Florence, Italy; Univ Florence, Dept Biol, Florence, Italy; CNR, Natl Inst Opt, Florence, Italy; Univ Florence, Dept Phys & Astron, Florence, Italy; Univ Florence, Dept Neurosci Psychol Drugs & Child Hlth, Florence, Italy; Univ Freiburg, Inst Expt Cardiovasc Med, Fac Med, Fribourg, Switzerland.
Abstract: Both genetic and non-genetic cardiac diseases can cause severe remodeling processes in the heart. Structural remodeling, such as collagen deposition (fibrosis) and cellular misalignment, can affect electrical conduction, introduce electromechanical dysfunctions and, eventually, lead to arrhythmia. Current predictive models of these functional alterations are based on non-integrated and low-resolution structural information. Placing this framework on a different order of magnitude is challenging due to the inefficacy of standard imaging methods in performing high resolution imaging in massive tissue. In this work, we describe a methodological framework that allows imaging of whole mouse hearts with micrometric resolution. The achievement of this goal has required a technological effort where advances in tissue transformation and imaging methods have been combined. First, we describe an optimized CLARITY protocol capable of transforming an intact heart into a nanoporous, hydrogel-hybridized, lipid-free form that allows high transparency and deep staining. Then, a fluorescence light-sheet microscope able to rapidly acquire images of a mesoscopic field of view (mm-scale) with the micron-scale resolution is described. Following the mesoSPIM project, the conceived microscope allows the reconstruction of the whole mouse heart with micrometric resolution in a single tomographic scan. We believe that this methodological framework will allow clarifying the involvement of the cytoarchitecture disarray in the electrical dysfunctions and pave the way for a comprehensive model that considers both the functional and structural data, thus enabling a unified investigation of the structural causes that lead to electrical and mechanical alterations after the tissue remodeling.
Journal/Review: JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume: (176) Pages from: e62795-1 to: e62795-13
More Information: This project has received fundings from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 952166 (REPAIR) , MUR under the FISR program, project FISR2019_00320 and Regione Toscana, Bando Ricerca Salute 2018, PERCARE project.KeyWords: Animals; Heart; Mice; Microscopy; Optical ImagingDOI: 10.3791/62795Citations: 4data from “WEB OF SCIENCE” (of Thomson Reuters) are update at: 2024-10-27References taken from IsiWeb of Knowledge: (subscribers only)Connecting to view paper tab on IsiWeb: Click hereConnecting to view citations from IsiWeb: Click here