Dmr with retina qimaging camera

Formalin solution (Anachemia, Montreal, QC, Canada) was used to fix the proximal sections (~10 mm) from each tibia for a duration of 48h. Thereafter, graded alcohol solutions were used for dehydration, xylene was used for clarification and methylmethacrylate (MMA) (Fisher Scientific Canada, Nepean, ON, Canada) was used for embedding process [35 (link)]. When the polymerization was completed, a microtome (Leica SM2500) setup was used to cut the blocks of the tibiae into 6 μm sections. Only the proximal sections were used in this study. To cover the 40–50% of the growth plate depth, the tibiae were cut along the longitudinal bone axis for 36 slides, six series of six slides, which contain two sections per slide. To facilitate the growth rate measurements, the first slide of each series (6 slides, 12 sections total) per proximal tibia were set aside from light. A microscope (Leica DMR with Retina Qimaging Camera) was used for slice observation while using 5x magnification for growth rate measurements.

Proximal sections (~10 mm) from each tibia were fixed for 48 h using the formalin solution (Anachemia, Montreal, QC, Canada). For dehydration and clarification, graded alcohol solutions and xylene were used respectively. Embedding process was performed using methylmethacrylate (MMA) (Fisher Scientific Canada, Nepean, ON, Canada)^{43 (link)}. After polymerization, tibial blocks were cut in 6 µm sections using a microtome (Leica SM2500). The longitudinal bone axis was used to cut the tibiae for 36 slides, six series of six slides, containing two sections per slide. For each tibia, the first slide of each series was set aside from light for growth rate measurements, for a total of 6 slides (i.e., 12 sections total). Microscopic observations were performed with a microscope (Leica DMR with Retina Qimaging Camera) using 2.5x magnification.