Slr based photo slit lamp imaging system

Mouse eyes were examined with a Topcon SL-D8Z slit lamp biomicroscope with a Nikon SLR-based Photo Slit Lamp imaging system following mydriasis with 1% Atropine Sulfate (Bausch & Lomb). Eyes, brains, and testes were collected at 8 weeks of age. Eyes and testes were fixed in 4% paraformaldehyde (PFA), paraffin embedded and H&E stained as previously described [5 (link)]. Brains were fixed at 4°C for 24 h in 4% PFA followed by 30% sucrose for 24-72 hrs. Brains were then sectioned at 30 μm on a sliding microtome (Leica) and stained with DAPI to label all nuclei. Immunostaining was done with TRA54 (B-Bridge) as a primary antibody and DyLight 488 goat anti-rat (Abcam) as a secondary antibody following the manufacturer’s recommendations. PNA staining was performed utilizing the Lectin PNA-Alexa-488 conjugate (Life Technologies) according to the manufacturer’s recommendations. Slides were DAPI stained according to the manufacturer’s recommendations (Life Technologies), mounted using Fluoromount-G (Southern Biotech), and imaged using a Nikon DS-Fi1 camera on a Nikon Eclipse 80i microscope using NIS-Elements software (Nikon).

Weights of WT (n = 8) and bs2 (n = 8) postnatal mice were measured and recorded in littermates from bs2/+ × bs2/+ crosses between P0.5 and 4 months of age. Age-matched bs2 (n = 4), AGPS-KOMP mice (n = 4), Agps-KOMP EIIa-Cre (n = 2) and control (n = 4) mice were X-ray imaged at 4 months of age. Exposures were recorded at a peak kilovoltage of 50 kVp and a charge of 0.50 mAs (milliampere seconds). The same mice were also evaluated with a Topcon SL-D8Z slit lamp biomicroscope with a Nikon SLR-based Photo Slit Lamp imaging system following mydriasis with 1% Atropine Sulfate (Bausch & Lomb). WT (n = 6) and bs2 (n = 6) testes weights were measured in age-matched pairs between 4 and 8 weeks of age. Significance for all measurements was calculated via two-tailed t-test (GraphPad), where P < 0.05 was considered significant.