Cell surface glycoproteins and glycans play critical roles in a range of biological functions and disease processes, from maintaining cellular structure and adhesion to controlling how cells send and receive exogenous signals in a complex environment. In the heart, for example, proper expression and glycosylation of ion channels are essential for propagating action potentials and proper contraction of the myocardium. Despite their critical roles in cardiac development, disease, and drug uptake, we do not yet have a detailed cell type- or chamber-resolved view of the cell surface glycoproteome or glycome of the adult human heart. Combining advanced strategies to isolate individual cardiac cell types with the recently developed CellSurfer Platform, which integrates a microscale Cell Surface Capture method for the identification of cell surface glycoproteins from small sample sizes, automated data processing workflows, and SurfaceGenie for cell-type specific marker prioritization, new views of the human heart are emerging. To date, we have identified >650 cell surface N-glycoproteins on primary human cardiac myocytes and fibroblasts, including proteins not previously described in these cell types and putative cell-type and chamber-specific markers. Additionally, a structure-based glycomics approach reveals >110 glycan structures in cardiac myocytes, which complement and inform glycoproteomic efforts. Overall, these data represent the first major step towards a comprehensive, cell-type, subtype, and chamber-resolved reference map of cell surface glycoproteins and glycans in the adult human heart and reveal new potential targets for immunophenotyping, in vivo imaging, drug delivery, and benchmarking cardiomyocytes derived from human pluripotent stem cells. Moreover, these data inform caveats regarding the use of explanted cardiac fibroblast models, reveal new molecular targets to study in the context of cardiac fibrosis and heart failure, and will promote studies aimed at gaining a better understanding of cross-talk among cardiac cell types in health and disease.