Registration-based 3D Light Sheet Fluorescence Microscopy and 2D histology image fusion tool for pathological specimen
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Background
Histological analysis traditionally relies on thin tissue sections, providing inherently two-dimensional (2D) information. However, this approach captures only a fraction of the entire sample and lacks the spatial context nec-essary for comprehensive tissue assessment. Recent advancements in multimodal imaging have introduced the fusion of histological data with three-dimensional (3D) imaging techniques, such as Light Sheet Fluorescence Microscopy (LSFM), to enhance tissue analysis by integrating complementary spatial information. A key challenge in this fusion process is the accurate alignment of corresponding structures across modalities, which is complicated by differences in resolution, sectioning-induced deformations, and varying imaging orientations. Existing solu-tions often require manual selection of image pairs or technical expertise, limiting accessibility to non-specialist users.
Methods
To address these limitations, we introduce LitSHi (Light Sheet meets Histology), a novel registration tool that enables the automated and precise align-ment of LSFM and histological images. LitSHi allows multimodal image fusion to be performed fully automatically, which significantly reduces the need for manual intervention.
Results
We evaluated LitSHi on testicular tumor specimens, demonstrating its ability to achieve enhanced structural correspondence between LSFM and histological images. The automated registration process significantly improved efficiency and alignment accuracy compared to traditional manual or semi-automated approaches.
Conclusion
LitSHi could improve digital pathology by optimizing multimodal tissue analysis and supporting future developments in computational pathology and AI-driven diagnostics.
