When imaging tissues, it is very important to minimize refracted index mismatches as it will cause imaging artifacts which increase with depth. Microscope objectives are designed to perform at specific refractive indices and are matched to common mediums such as air, water, glycerol, or oil. Some objectives will have correction collars so they can be used with many different refractive indices. For example, ideally an objective that is matched to a refractive index of 1.33 for water will be dipped in water and used to image a tissue that has been cleared with an aqueous clearing technique where the refractive index is also 1.33. When imaging the tissue, try to always match refractive indices for optimal imaging depth and image quality.
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