IBEX: A versatile multiplex optical imaging approach for deep phenotyping and spatial analysis of cells in complex...

Area of Science:

• Biotechnology
• Molecular Biology
• Cell Biology

Background:

• Understanding mammalian tissue complexity and cell-cell interactions is crucial for organ homeostasis and disease research.
• Current methods like single-cell genomics lack spatial context, and traditional immunofluorescence is limited in molecular feature detection.
• Existing advanced imaging technologies have limitations hindering widespread adoption.

Purpose of the Study:

• To develop a method overcoming limitations in high-content multiplex tissue imaging.
• To enable high-resolution imaging of a large number of molecular parameters in the same tissue section.
• To provide a versatile and adaptable imaging solution for diverse research applications.

Main Methods:

• Iterative Bleaching extends Multiplexity (IBEX) employs iterative staining and chemical bleaching.
• The method allows high-resolution imaging of over 65 parameters without physical tissue degradation.
• IBEX is compatible with conventional microscopes, various antibodies, and amplified or oligonucleotide-based staining.

Main Results:

• IBEX successfully images >65 parameters in the same tissue section with high resolution.
• The method is adaptable to different staining techniques and antibody types, including user-generated ones.
• An open-source platform for automated image registration facilitates data processing.

Conclusions:

• IBEX overcomes major impediments in multiplex tissue imaging, offering a powerful tool for biological research.
• This technology can be readily integrated into existing laboratory workflows for high-content imaging.
• IBEX enables detailed visualization of complex cellular landscapes in diverse organs and tissues, aiding in understanding homeostasis and disease.