Resolving cell state in iPSC-derived human neural samples with multiplexed fluorescence imaging.

Area of Science:

• Neuroscience
• Stem Cell Biology
• Biotechnology

Background:

• Human induced pluripotent stem cell-derived (iPSC) neural cultures are valuable models for neurological diseases like Alzheimer's and Autism Spectrum Disorder.
• Understanding cell state evolution and protein localization in 3D and 2D cultures is crucial for disease research.

Purpose of the Study:

• To develop and apply a novel multiplexed imaging technique for analyzing complex iPSC-derived neural cultures.
• To facilitate in situ characterization of spatial-temporal protein expression in neural differentiation and disease models.

Main Methods:

• Application of PRobe-based Imaging for Sequential Multiplexing (PRISM) for rapid, facile exchange of imaging probes.
• Multiplexed imaging of iPSC-derived cortical and motor neuron cultures using over ten protein targets.
• Analysis of cell differentiation, composition, and functional markers in complex neural cultures.

Main Results:

• PRISM enabled detailed analysis of protein expression and localization in iPSC-derived neural cultures.
• The method successfully characterized cell differentiation and marker expression relevant to psychiatric and neurodegenerative diseases.
• Demonstrated the potential for automation and scalability to numerous protein targets and samples.

Conclusions:

• PRISM is an effective approach for analyzing complex stem-cell derived neural cultures.
• This technique enhances understanding of neural cell differentiation and disease phenotypes.
• The method's scalability and automation potential offer significant advantages for future research.