In vivo clonal analysis of cardiomyocytes

Abstract

The development of the CreER/LoxP system has enabled temporal control and cell type specificity of gene activation or repression. A common application of this system involves lineage tracing and examining the proliferative capacity of cells of interest through clonal analysis. Here, we describe a method of performing 2- and 3-dimensional clonal analysis of cardiomyocytes (CMs) using the Rainbow reporter mouse model. We outline the process of using the Cell Counter plug-in tool in ImageJ to quantify the number of clones as well as the number of cells within each clone. For 3-dimensional analysis, we describe the tissue clearing technique, CLARITY, in conjunction with light-sheet imaging to obtain digital slices of the whole heart that can be reconstructed and clone volumes quantified using the Imaris software.