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Developing a novel hyperspectral imaging cryomacrotome for whole body fluorescence imaging
Boyu Meng1, Brook K Byrd1, Dennis J Wirth1
1Thayer School of Engineering, Dartmouth College, Hanover, NH 03755.
Summary
We developed a hyperspectral imaging whole body cryo-macrotome for high-resolution 3-D fluorescence imaging in whole animals. This technology allows precise tracking of cells, metastasis, and drug distribution by separating multiple fluorescent signals.
Area of Science:
- Biomedical Imaging
- Optical Spectroscopy
- Preclinical Research
Background:
- Whole-body fluorescence imaging is crucial for tracking biological processes like cell migration and drug distribution.
- Existing methods often lack the resolution or spectral capabilities for complex analyses in whole animals.
- Accurate quantification and separation of multiple fluorophores and autofluorescence are significant challenges.
Purpose of the Study:
- To develop and validate a novel hyperspectral imaging whole body cryo-macrotome for high-resolution 3-D fluorescence volume acquisition.
- To enable spectral decoupling of multiple fluorophores and autofluorescence within large biological specimens.
- To demonstrate the system's utility in preclinical research, specifically in a rodent glioma xenograft model.
Main Methods:
- Development of a custom-built hyperspectral imaging whole body cryo-macrotome.
- Acquisition of 3-D fluorescence volumes at high resolution from whole animal specimens.
- Utilizing hyperspectral data to obtain full emission spectra at each voxel.
- Applying spectral unmixing algorithms for fluorophore and autofluorescence decoupling.
Main Results:
- Successful acquisition of high-resolution 3-D fluorescence volumes from whole rodent specimens.
- Demonstrated ability to spectrally decouple multiple fluorophores and endogenous autofluorescence.
- Presented characteristic tissue spectra and successful spectral fitting in a rodent glioma xenograft model.
- High spatial resolution enables detailed visualization of biological processes.
Conclusions:
- The developed hyperspectral imaging whole body cryo-macrotome is a powerful tool for advanced preclinical research.
- This technology significantly enhances the ability to track labeled cells, metastatic spread, and drug bio-distribution in vivo.
- Spectral decoupling capabilities provide unprecedented accuracy in quantitative fluorescence imaging of whole organisms.
