Dp58 camera

We used 6-week-old male Sprague Dawley rats. Rats were euthanized with CO₂ asphyxiation, and eyeballs were removed. Next, we enucleated lenses in PBS with using a dissection microscope. Enucleated lens were cultured in 2 mL serum-free M199 medium (Sigma) containing 0.1% BSA [35 (link)]. MMS (Nakalai tesque) or Bleomycin (Tokyo chemical industry) were added to the culture medium at final concentrations of 1 mM and 800 μg/mL, respectively. Culture medium was changed once daily, and fresh drug was added to the medium every time it was changed. All lenses were maintained at 37°C in a humidified incubator with 95% room air and 5% CO₂. On the last day of culture, we photographed lenses with a microscope using a DP58 camera (Olympus) attached to an SZX12 stereomicroscope in a 35 mm dish containing 7 mL PBS. All experiments were approved by the Animal Research Committee of the University of Fukui (Approval number: 28091) and were conducted in accordance with the University of Fukui Animal Care and Use Regulations and Association for Research in Vision and Ophthalmology Statement for the Use of Animals in Ophthalmic and Vision Research. The findings of the study were reported in accordance with ARRIVE guidelines.

Images were captured in the dark using a SZX12 stereomicroscope combined with a DP58 camera (Olympus). At this time, the lens was placed in a 35 mm dish containing 7 mL PBS. The degree of opacity was measured as brightness (~0–255) using ImageJ, and a weighted average was calculated.

Photographs of the lens were taken in the darkroom using an SZX12 stereomicroscope with a DP58 camera (Olympus) attached, as previously shown^{54 (link)}. Photographs were taken in 35 mm petri dishes containing 7 mL of PBS. A weighted average was calculated from the brightness (0–255) of the cortex area of the lens that was opacified by incubation with galactose, and the weighted average was again calculated from the brightness of the same area in the lens after addition of the inhibitor and further incubation^{30 (link)}. The change in opacity was calculated by subtracting the value after the addition of the inhibitor from the value before the addition of the inhibitor.