Figure 2. Scattered light reduction and data fusion with electronic confocal slit detection.

(a) Left- and right-sided illuminations of a single (sagittal) plane 50 μm inside the Drosophila embryo expressing His2Av-RFP1 is shown for widefield and eCSD detection. The decrease in image quality away from the illumination side is clearly visible in widefield detection, while eCSD removes these blurred regions. (b) Intensity profiles across the embryo for indicated regions in (a). Intensity values close to and away from the illumination side are of equal magnitude for widefield detection (brown and green lines) preventing a direct fusion (addition) of images. eCSD profiles decay away from the illumination side as scattered photon are blocked. Please also note the removal of the scattered photon haze outside the embryo. (c) eCSD detection with simultaneous illumination yields superior image quality compared to the established imaging procedure of acquiring two sequential widefield images followed by a sigmoidal fusion step. Simultaneous widefield and sigmoidal fused eCSD images are shown for completeness. Magnified views of all four imaging procedures are shown on the right. Scale bars are 50 μm in both (a,c) except inserts (30 μm). For all illumination and detection schemes optical (xy) sections 50 μm deep inside an embryo at germ band retraction stage were imaged.