A mechanical scanning acoustic reflection microscope was applied to living cells (e.g., osteoblasts) to observe their undisguised shapes and to evaluate their adhesive conditions at a substrate interface. A conditioned medium was collected from a bone-metastatic breast cancer cell line, MDA-MB-231, and cultured with an immature osteoblast cell line, MC3T3-E1. To characterize the cellular adhesion, MC3T3-E1 osteoblasts were cultured with or without MDA-MB-231 conditioned medium for 2 days, then assayed with the scanning acoustic reflection microscope. At 600 MHz the scanning acoustic reflection microscope clearly indicated that MC3T3-E1 cells cultured with MDA-MB-231 conditioned medium had both an abnormal shape and poor adhesion at the substrate interface. The results are compared with those obtained with laser scanning confocal microscopy and are supported by a simple multilayer model.