Imaging transparent intact cardiac tissue with single-cell resolution

Abstract

We developed a new method, SUT (Scheme Update on tissue Transparency), to view cardiac microstructures and unveil the molecular changes underlying cardiac diseases. SUT is an effective method to clear whole-hearts from different species. Over the course of 4 - 6 days we obtained transparent whole-layer left ventricular tissues from mice with only an approximate 1% protein loss. In addition, EAL (Electrophoretic Antibody Labeling) was used to achieve fast antibody labeling by electric force, which significantly reduced antibody incubation time from days to hours. SUT, together with EAL and modern imaging techniques, were successfully used to visualize three-dimensional spatial distribution of various molecules in cardiac tissue. We also observed changes in the number and phenotypes of fibroblasts during post-myocardial infarction in a stereoscopic pattern. We believe that our technique opens a new avenue to explore the mechanisms underlying cardiac diseases.

Biomedical Applications of Tissue Clearing and Three-Dimensional Imaging in Health and Disease

“…The whole mouse heart can be cleared with CUBIC allowing 3D vasculature image ( Nehrhoff et al., 2017 ) ( Figure 4 ). A combination of CUBIC and CLARITY, named SUT (Scheme Update on tissue Transparency), has been used to evaluate spatial distribution and phenotype of fibroblasts in mice left ventricles in a post-myocardial infarction ( Wang et al., 2018 ). Additionally, a modified CLARITY method that skips the electrophoretic step named SCM (Simplified CLARITY method) has been applied to mouse heart to study proteins distribution ( Sung et al., 2016 ).…”

Tissue clearing and imaging methods for cardiovascular development

“…The main disadvantage is still a long clearing period for larger samples. This significant limitation was recently overcome with a modification named SUT (Wang et al, 2018).…”

Advances in Ex Situ Tissue Optical Clearing

“…Published by Springer Nature. e,i) Reproduced with permission . Copyright 2018, The Optical Society.…”

“…A major drawback of heart clearing by CUBIC is the incubation time, starting from 5 days for embryonic hearts, through 7–14 days for the whole embryos, and finally lasting 30 days for the adult hearts . This significant limitation was recently overcome by Wang et al., who described SUT (Scheme Update on tissue Transparency). In brief, SUT combines selected chemicals used in CLARITY/PACT methods (8% SDS) and CUBIC (Urea and Triton X‐100).…”