Comparisons were made of the cytoplasmic textures in electron microscope images of nuclear fiber cells from a variety of human and animal lenses. The goals were to establish the optimal conditions for quantifying the textural features and for relating the extent of roughness with the observed extent of nuclear opacification. Freshly cut Vibratome sections were fixed and processed for thin-section electron microscopy. Normal human donor lenses, human age-related cataracts from surgery, and rat, guinea pig, and canine lenses were analyzed using density linescans, Fourier transforms, and autocorrelation analysis. Normal and control lenses were compared to lenses with varying degrees of scattering including fully opaque nuclear cataract. Images were recorded at 21,000 x, giving structural information in the critical range of 2-300 nm. Human normal and nuclear cataractous lens cytoplasm produce Fourier transforms with relatively high intensity in the range 10-50 nm (equivalent spacing) and relatively low intensity greater than 100 nm. This is consistent with the smooth image appearance, linescans with small fluctuations and autocorrelation functions indicating that the images are nearly homogeneous. Images of the transparent animal lenses were very smooth and produced Fourier transforms that showed less intensity in the range 10-50 nm and less intensity greater than 100 nm compared to the human lenses. Animal lenses with progressively enhanced light scattering showed a strong correlation between increased textural roughness and increased Fourier intensity greater than 100 nm. These analytical image analysis techniques readily documented the wide range of cytoplasmic textural variations in human and animal lenses and cataracts. Consistent comparisons were possible only when well-preserved tissues were examined with high-resolution images. The cytoplasm with the greatest roughness correlated with the greatest light scattering suggests that redistribution and/or loss of cytoplasmic proteins contribute to cataract formation.

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http://dx.doi.org/10.1006/exer.2001.1166DOI Listing

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