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Image Processing

Fluorescence Stitching

Building seamless mosaics from fluorescence tiles

Definition
The process of seamlessly joining overlapping fluorescence image tiles into a single large mosaic while correcting for vignetting, intensity fall-off, and sub-pixel misalignment so that analysis algorithms operate uniformly across tile boundaries.
Flat-Field Correction
Eliminates illumination non-uniformity
Sub-Pixel Alignment
Tiles aligned with fraction-of-a-pixel precision

Stitching Challenges

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 engine corrects all of these artifacts, producing images where cell detection algorithms see no tile boundaries.

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