Fluorescence stitching is more demanding than brightfield because:
- Vignetting: illumination intensity falls off at tile edges, creating visible seams if uncorrected
- Photobleaching: overlapping regions may have reduced signal from double exposure
- Background variation: autofluorescence varies across the tissue, complicating intensity normalization
PanoBrain's stitching corrects vignetting and normalizes intensity across tile overlaps, so tile boundaries do not show in the images that cell detection runs on. Stitching cannot restore signal lost to photobleaching or remove autofluorescence from the tissue.