Pocket CLARITY enables distortion-mitigated cardiac microstructural tissue characterization of large-scale specimens

Joan J H Kim; Shestruma Parajuli; Aman Sinha; Mohammed Mahamdeh; Maaike van den Boomen; Jaume Coll-Font; Lily Shi Chen; Yiling Fan; Robert A Eder; Kellie Phipps; Shiaulou Yuan; Christopher Nguyen

doi:10.3389/fcvm.2022.1037500

Pocket CLARITY enables distortion-mitigated cardiac microstructural tissue characterization of large-scale specimens

Front Cardiovasc Med. 2022 Nov 14:9:1037500. doi: 10.3389/fcvm.2022.1037500. eCollection 2022.

Authors

Joan J H Kim¹, Shestruma Parajuli¹, Aman Sinha¹, Mohammed Mahamdeh^{1

2}, Maaike van den Boomen^{1

2

3

4}, Jaume Coll-Font^{1

2

4}, Lily Shi Chen¹, Yiling Fan^{1

2

5}, Robert A Eder¹, Kellie Phipps¹, Shiaulou Yuan^{1

2

4}, Christopher Nguyen^{1

2

4

6

7}

Affiliations

¹ Cardiovascular Research Center, Massachusetts General Hospital, Charlestown, MA, United States.
² A.A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Boston, MA, United States.
³ Department of Radiology, University Medical Center Groningen, University of Groningen, Groningen, Netherlands.
⁴ Department of Medicine, Harvard Medical School, Boston, MA, United States.
⁵ Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, United States.
⁶ Division of Health Science Technology, Harvard-Massachusetts Institute of Technology, Cambridge, MA, United States.
⁷ Cardiovascular Innovation Research Center, Heart, Vascular, and Thoracic Institute, Cleveland Clinic, Cleveland, OH, United States.

Abstract

Molecular phenotyping by imaging of intact tissues has been used to reveal 3D molecular and structural coherence in tissue samples using tissue clearing techniques. However, clearing and imaging of cardiac tissue remains challenging for large-scale (>100 mm³) specimens due to sample distortion. Thus, directly assessing tissue microstructural geometric properties confounded by distortion such as cardiac helicity has been limited. To combat sample distortion, we developed a passive CLARITY technique (Pocket CLARITY) that utilizes a permeable cotton mesh pocket to encapsulate the sample to clear large-scale cardiac swine samples with minimal tissue deformation and protein loss. Combined with light sheet auto-fluorescent and scattering microscopy, Pocket CLARITY enabled the characterization of myocardial microstructural helicity of cardiac tissue from control, heart failure, and myocardial infarction in swine. Pocket CLARITY revealed with high fidelity that transmural microstructural helicity of the heart is significantly depressed in cardiovascular disease (CVD), thereby revealing new insights at the tissue level associated with impaired cardiac function.

Keywords: CLARITY; distortion; helicity; lightsheet microscopy; microstructure.

Abstract

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