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Updated: Jun 8, 2026

A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
Published on: March 30, 2020
Imaging the bio-distribution of fluorescent probes using multispectral epi-illumination cryoslicing imaging
Athanasios Sarantopoulos1, George Themelis, Vasilis Ntziachristos
1Chair for Biological Imaging, Technische Universität München, Munich, Germany.
Purpose:
The increasing availability of fluorescent probes for in vivo optical imaging enables the interrogation of complex biological processes in small animals serving as models for human-like tissue function and disease. However, the validation of probe bio-distribution during their development or the study of different disease models, in support of in vivo imaging studies, is not straightforward.
Procedures:
The imaging system developed consists of a customized multispectral planar imager that has been adapted on a commercial cryomicrotome and provides a powerful modality for ex vivo imaging of small animals.
Results:
The ability to capture 3D anatomical (color) and fluorescence volumetric distributions of multiple fluorescent markers in high resolution is showcased.
Conclusions:
Serving both as a method for accurately imaging the bio-distribution of multiple fluorescent agents inside organisms and as a modality for the validation of non-invasive methods, multispectral cryoslicing imaging offers useful insights to ex vivo optical imaging of molecular probes.

