Childhood cancer survivors (CCSs) face lifelong side effects related to their treatment with chemotherapy. Anthracycline agents, such as doxorubicin (DOX), are important in the treatment of childhood cancers but are associated with cardiotoxicity. Cardiac toxicities represent a significant source of chronic disability that cancer survivors face; despite this, the chronic cardiotoxicity phenotype and how it relates to acute toxicity remains poorly defined., To address this critical knowledge gap, we studied the acute effect of DOX on murine cardiac non-myocytes in vivo. Determination of the acute cellular effects of DOX on non-myocytes, a cell pool with finite replicative capacity, provides a basis for understanding the pathogenesis of the chronic heart disease that CCSs face. To investigate the acute cellular effects of DOX, we present scRNAseq data from homeostatic cardiac non-myocytes and compare it to pre-existing datasets as well as a novel CyTOF datasets. SCANPY, a python-based single-cell analysis, was used to assess the heterogeneity of cells detected in scRNAseq and CyTOF. To further assist in CyTOF data annotation, joint analyses of scRNAseq and CyTOF data using an artificial neural network known as sparse autoencoder for clustering, imputation, and embedding (SAUCIE) are performed. Lastly, the panel is tested on a mouse model of acute DOX exposure at two-time points (24 and 72 hours) after the last dose of doxorubicin and examined with joint clustering. In sum, we report the first-ever CyTOF study of cardiac non-myocytes and characterize the effect of acute DOX exposure with scRNAseq and CyTOF.