Multielement Z-tag imaging by X-ray fluorescence microscopy for next-generation multiplex imaging.

Area of Science:

• Biophysics
• Molecular Imaging
• Biotechnology

Background:

• Established imaging methods face limitations in simultaneously achieving high multiplexing, speed, and resolution.
• X-ray imaging offers potential for deep tissue penetration and atomic-level detail.

Purpose of the Study:

• To develop a rapid, highly multiplexed, and nondestructive imaging technique for tissue analysis.
• To overcome the physical constraints of current bioimaging modalities.

Main Methods:

• Development of multielement Z-tag X-ray fluorescence (MEZ-XRF).
• Integration of MEZ-XRF with signal amplification by exchange reaction (SABER)-amplified Z-tag reagents for enhanced speed and sensitivity.
• Demonstration of parallel imaging of 20 Z-tag or SABER Z-tag reagents.

Main Results:

• Achieved subcellular resolution imaging in cell lines and human tissues.
• Demonstrated parallel imaging of up to 20 reagents simultaneously.
• Benchmarked MEZ-XRF against imaging mass cytometry.
• Showcased nondestructive, multiscale repeat imaging capabilities, from rapid overview scans to sensitive detection of low-abundance markers.

Conclusions:

• MEZ-XRF provides a powerful tool for highly multiplexed bioimaging across diverse biological scales.
• The combination of MEZ-XRF and SABER Z-tags offers high sensitivity and speed for advanced tissue analysis.
• This technique enables nondestructive, multiscale imaging, advancing tissue analysis and biomarker discovery.