Comparison of different methods applied to an entire specimen of Dikerogammarus villosus stored in alcohol. All images are fused from several focal planes. (a)–(c) Macrographs. (a) “Normal” macrograph, with optimized uniform light. Bubbles are evident on the black velvet, which was used as background. Due to reflections, individual elements of the limbs are difficult to distinguish (arrow). (b) Macrograph under polarized light. Bubbles are not apparent. Borders between individual elements of the limbs more conspicuous (arrow). (c) Macrofluorescence. Borders between individual limb elements are well apparent due to the color contrast (arrow). (d) Composite fluorescence image [4]. Details well contrasted, also the individual limb elements (arrow).
