Abstract Background/Objective: Osteoporosis is mainly characterized by a deterioration of microstructure and a loss of biochemical components in bone tissues. Developing an imaging technique for measuring bone tissue microstructure and the biochemical components is of great significance for the early diagnosis of osteoporosis. Photoacoustic microscopy (PAM) has the advantage of high optical resolution and the potential to diagnose osteoporosis. In this study, we investigated the feasibility of the photoacoustic microscopy (PAM) technique for bone tissue imaging, and the deterioration of microstructure and biochemical components in cancellous bone was characterized by the PAM. Statement of Contribution/Methods: We performed the optical-resolution photoacoustic microscopy (OR-PAM) for bone tissue imaging and the trabecular microstructure and hydroxyapatite (HAP) were degraded by immersion in JYBL-I solution. The PAM imaging method was developed for the measurement of the surface and subsurface of cancellous bone with a high resolution. Specifically, a 532 nm pulse laser was used to excite the PA signal from the bone. The PA signal sampling frequency was 80 MHz. A motor rotated a 15 MHz central frequency transducer to receive 1000 × 1000 × 250 points data. The envelope of the signal was obtained using the Hilbert transform for reconstruction. Then, the JYBL-I solution was used to reduce the HAP component in the bone. The PAM imaging was performed after different immersion times, (i.e., at 0, 5, and 10 mins). In the PAM measurement of the cancellous bone, the imaging area was a cylinder with an 8 mm diameter and an 8 μm/pixel resolution. Results/Discussion: The results showed that the trabecular microstructure could be imaged with a relatively high quality using the PAM technique. With the different extent of HAP degradation by immersion in JYBL-I solution, some trabecular bone disappeared corresponding with PA signals decreased significantly in amplitude. Conclusion: These results indicate that the PA technique has potential application in the characterization of bone microstructure and biochemical components with a high resolution.