Comparison of matched pairs of WSI images using DIRI side illuminator and traditional darkfield substage condenser.

<p>Three WSI images using DIRI system (6A, 6C) are compared with traditional darkfield substage condenser images (6B, 6D). Each pair of photos (6A–B and 6C–D) depict the same region of a slide. The images using the DIRI system initially do not appear as robust as those taken with a darkfield substage condenser. Further examination of the images reveals that the images obtained with the DIRI system are more accurate representations of the cells and their processes. The substage darkfield condenser typically results in excessive light scatter. Differences in brightness and color of the individual cells with the DIRI system vs. the substage darkfield condenser are due in part to the difference in brightness of the illuminators and the spectral features of the illumination sources. (6E) shows the same region depicted in 6C–D, but is based upon a brightfield image that was then inverted to provide a “pseudo-darkfield” effect. Either darkfield method provides much better detail of the microscopic field than the inverted image in 6E.