Three-dimensional imaging enhances delineation of cardiac anatomy and function. Currently, three-dimensional echocardiography involves complex rendering techniques to imply depth in a given image. Also, display of the final images on a video monitor neutralizes the volume information of the object. A hologram is a true, three-dimensional replica of the original object and does not entail complex data processing. Holography has not been used previously in cardiovascular imaging. In this experiment, 11 excised mammalian whole hearts and five isolated left ventricles were imaged in a water bath. Parallel, tomographic, echocardiographic images comprised the data source from which holograms were obtained by the technique of multiplexed holography. Holograms were mounted on a holographic film and viewed in a special viewing box. High-quality holograms were obtained from every data set. Excellent gray-scale discrimination allowed crisp visualization of cardiac structures and simulated diseases. Electronic sectioning and various projections optimized viewing of the three-dimensional anatomy in surgical orientations. Thus holograms can be produced from tomographic cardiac ultrasound data and could open a new avenue in three-dimensional echocardiography.