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. 2021 Aug 15;13(8):8921–8937.

Figure 4.

Figure 4

CA4+ labeled cartilage imaging after decalcification. A. 2D images of cartilage at the patellar groove level. The unfixed tissues have lost cartilage and bone signal after decalcification. Bone is not visible in the no-CA4+ labeled tissues. In contrast, the cartilage signal at the patellar groove and condyle of NBF fixed tissues showed visible cartilage structure. No signal is visualized in the subchondral bone except for residual trabecular bone in the 24- and 30-hour CA4+ labeled tissues. Scale bars = 5 mm. B. Micro3D image demonstrates clearly defined patellar groove cartilage and condyle cartilage in the NBF fixed cartilage after decalcification. Scale bars = 1 mm. C. Comparison of cartilage density for fresh CA4+ labeled, PBS soaked, and decalcified NBF fixed tissues. Specimens soaked in PBS for 5 days did not cause signal loss compared to fresh-labeled cartilage. Decalcification caused approximately a 20% decrease in density compared to fresh labeled and PBS soaked tissues. **P<0.01, ***P<0.001, ****P<0.0001. D. Quantification of cartilage volume showed no statistical differences after decalcification compared to fresh labeled, PBS soaked for 5 days.