The cartilage-conduction pathway was recently proposed as a third auditory pathway; however, middle-ear vibrations have not yet been investigated in vivo. We aimed to measure the ossicles and bone vibration upon cartilage-conduction stimulation with a non-contact laser Doppler vibrometer. We recruited adult patients with normal ear structures who underwent cochlear implant surgery at our hospital between April 2020 and December 2022. For sound input, a cartilage-conduction transducer, custom-made by RION Corporation (Tokyo, Japan), was fixed to the surface of the tragus and connected to an audiometer to regulate the output. A posterior tympanotomy was performed and a laser beam was directed through the cavity to measure the vibration of the ossicles, cochlear promontory, and posterior wall of the external auditory canal. Five participants (three men, mean age: 56.4 years) were included. The mean hearing loss on the operative side was 96.3 dB HL in one patient, and that of the other patients was off-scale. The vibrations were measured at a sound input of 1 kHz and 60 dB. We observed vibrations of all three structures, demonstrating the existence of cartilage-conduction pathways in vivo. These results may help uncover the mechanisms of the cartilage-conduction pathway in the future.