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Fluorescence-based cell sorting instruments are capable of classifying sperm for desired characteristics such as gender. It is not surprising that, as these techniques are approaching clinical use, closer scrutiny has revealed that the sorting process introduces a signification increase in oxidative stress in addition to an undesired selection bias due to fragmented DNA. Although some of the damage can be attributed to rough mechanical handling, from studies on adherent cells, it is well known that these defects are also associated with the intrinsic toxicity of the fluorescence labels. In effect, fluorescence-based markers reduce viability, motivating the use of intrinsic contrast methods. Here we present recent progress on evaluating reproductive outcomes using Spatial Light Interference Microscopy (SLIM) for high sensitivity phase imaging. We compared our label-free markers to existing staining techniques. We show how the compatibility of our system with fluorescence/color image acquisition allows our techniques to be used concurrently rather than as an alternative, to conventional methods.
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