Figure 5. Effect of deconvolution.

Cropped and zoomed-in images and intensity profiles on the surface texture of Croton hirtus and Agropyron repens, showing differences in the signal-to-noise ratio of each technique and the impact of blind deconvolution. CF = confocal; 2P = two-photon; WF = apotome; SNR = signal-to-noise ratio; Decon.  = deconvolved. SNR calculated as follows: x−y/x+y where x = the average of six maximum pixel intensity values from the peaks of the line profile, and y = the average of six minimum pixel intensity values from the troughs of the line profile (both areas randomly selected). In the case of C. hirtus, line profiles were traced on a single full Croton structure on the same pollen grain. In the case of A. repens, line profiles were traced on randomly selected regions of A. repens specimens. Data from widefield and apotome microscopy is from a single pollen grain. Data from confocal (405 nm and 561 nm) and two-photon microscopy is from a single pollen grain, but not the same specimen analysed used widefield and apotome microscopy. Intensity profiles were not drawn on precisely the same pixels in X-Y positions owing to the movement of pollen grains in the mounting media during the movement of slides between microscopes. Detailed images measure 5 µm in the X-Y direction and are displayed in linear intensity profiles.