Ccd camera

Immunoblot analyses were performed as previously described^{25 (link)}. Anti-Col1A, anti-CTGF, anti-Collagen type IV, anti- Apoptosis-Associated Speck-Like Protein Containing CARD (ASC), anti-CASP1, anti-CASP1p10, anti-NLRP3, anti-Actin antibodies (Abcam, USA) were used in combination with appropriate peroxidase-conjugated secondary antibodies. Tubulin or Actin were used as a loading control (Sigma, USA). Bands were visualized with the enhanced chemiluminescence reagent (Thermo, USA) and digitized using a CCD camera (UVP, USA). Densitometric analysis and quantification were performed using ImageJ software (NIH, USA).

Lenses were imaged using a charge coupled device (CCD) camera (UVP, Cambridge, UK) with Synoptics software (Synoptics, Cambridge, UK) at the experimental start point (day 0) and at 24 h intervals throughout experiments. Brightfield illumination was used, with a black and white grid placed beneath lenses to give an assessment of visual quality and therefore lens clarity. Visual quality was quantified from these images by measuring standard deviation values of grey scale values obtained from the grid beneath the lens. Values of standard deviation of clear lenses are high whereas when the lens becomes more opaque, the grid becomes less defined/homogenous and the standard deviation values decrease. A background for each image was achieved by selecting a region of the grid adjacent to the lens. This region exhibits the greatest standard deviation and best visual quality and thus a decrease was calculated relative to these values. Image analysis was performed with Image-Pro Premier (MediaCybernetics Inc., MD, USA) analysis software.