UVA Hyperspectral Light-Sheet Microscopy for Volumetric Metabolic Imaging: Application to Preimplantation Embryo...

Area of Science:

• Cellular and Molecular Biology
• Biophysics
• Microscopy and Imaging Technologies

Background:

• Cellular metabolism is crucial for cell energetics, growth, regeneration, and homeostasis.
• Spatially mapping metabolic heterogeneity is vital for understanding cell and tissue health.
• Current imaging techniques for endogenous metabolic cofactors like NAD(P)H and FAD have limitations for clinical applications.

Purpose of the Study:

• To develop a noninvasive, high-resolution 3D imaging method for mapping cellular metabolism.
• To utilize phasor-based hyperspectral light-sheet microscopy with a single UVA excitation wavelength.
• To enable real-time assessment of metabolic activity for clinical translation.

Main Methods:

• Developed a phasor-based hyperspectral light-sheet (HS-LS) microscopy system.
• Employed a single UVA excitation wavelength (375 nm) to simultaneously excite NAD(P)H and FAD autofluorescence.
• Utilized hardware-based spectral phasor analysis for quantifying relative contributions of NAD(P)H and FAD.

Main Results:

• Demonstrated simultaneous excitation and quantification of NAD(P)H and FAD autofluorescence using a single UVA wavelength.
• Captured dynamic changes in metabolic activity during preimplantation embryo development.
• Generated volumetric maps of metabolic activity to delineate developmental changes.

Conclusions:

• The developed HS-LS microscopy approach enables noninvasive 3D mapping of cellular metabolism.
• This technique overcomes limitations of existing methods, such as multiple excitation wavelengths or complex postprocessing.
• The findings pave the way for clinical translation, including in situ, noninvasive assessment of embryo viability and tissue health.