Multiplexed spatial transcriptomics methods and the application of expansion microscopy

Area of Science:

• Molecular Biology
• Genomics
• Biotechnology

Background:

• Spatial transcriptomics revolutionizes cellular organization studies.
• Numerous innovative transcriptomics methods have emerged, including in situ sequencing (ISS), in situ hybridization (ISH), and next-generation sequencing (NGS)-based region capture.
• A key challenge is limited RNA detection due to optical crowding.

Purpose of the Study:

• To review ISS, ISH, and NGS-based spatial transcriptomic protocols.
• To explore how expansion microscopy (ExM) enhances these techniques.
• To highlight the broader implications of transcriptomics in biology and medicine.

Main Methods:

• Review of existing literature on spatial transcriptomics techniques (ISS, ISH, NGS).
• Discussion of expansion microscopy (ExM) principles and applications.
• Integration of ExM with spatial transcriptomics protocols.

Main Results:

• ExM overcomes optical crowding and diffraction limits for improved RNA visualization.
• Combining ExM with ISS, ISH, and NGS enhances RNA detection sensitivity and spatial resolution.
• Spatial transcriptomics provides insights into gene expression, cell types, and disease mechanisms.

Conclusions:

• Expansion microscopy significantly improves spatial transcriptomics performance.
• These enhanced techniques offer deeper understanding of cellular organization and disease.
• Spatial transcriptomics holds vast potential for biological and medical research breakthroughs.