Specimens were fixed in 4% paraformaldehyde for 24 h for hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) of the limbs. P7 mice required decalcification for 24 h with 10% EDTA (pH 7.4) solution, and 4 weeks- and 4 months-old mice required decalcification for at least 5 days. After decalcification, dehydration, and paraffin infiltration were performed using an automated tissue processor (TP1020, Leica). Embedded tissues were cut to 6-µm-thick sections with a rotary microtome (RM2145, Leica). For IHC, the section slides were subjected to antigen retrieval in citrate buffer for 10 min at 90 °C. Primary antibodies to the following antigens were used: PADI2 (Proteintech), RUNX2 (MBL Life science), and Collagen type I (COL1) (sc-59772; Santa Cruz Biotechnology, Inc). All the stained images were acquired using a DP72 digital microimaging camera (Olympus) under a BX51 microscope (Olympus).
Dp72 digital microimaging camera
Manufactured by Olympus
The DP72 digital microimaging camera from Olympus is a high-performance camera designed for microscopy applications. It features a 12.8-megapixel CCD sensor and can capture images with a resolution of up to 4140 x 3096 pixels. The camera supports a range of image file formats, including TIFF, JPEG, and BMP. It is compatible with a variety of microscope systems and can be integrated into digital imaging workflows.
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Lab products found in correlation
2 protocols using dp72 digital microimaging camera
1
Histological Analysis of Mouse Limb Development
2
Histochemical Analysis of Murine Skeletal Development
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For hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC) of tissue sections, P21 mice were fixed in 4% paraformaldehyde for 24 h. Paraffinization of a dissected tibia was conducted (TP1020, Leica) after decalcification with 10% EDTA (pH 7.4). Detailed procedures were performed as described previously (Kim B. S. et al., 2021 (link)). IHC using antibodies against collagen Type X (COL10; 234,196, Millipore‐Sigma), proliferating cell nuclear antigen (PCNA; sc‐56, Santa Cruz Biotechnology), SOX9 (sc‐20095, Santa Cruz Biotechnology), MMP9 (sc6840, Santa Cruz Biotechnology), MMP13 (ab39012, Abcam), and nuclear factor‐κB (NF‐κB; Santa Cruz Biotechnology) was conducted after antigen retrieval. All stained tissue images were acquired with a DP72 digital microimaging camera (Olympus) under a BX51 microscope (Olympus).
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