Expansion and Light-Sheet Microscopy for Nanoscale 3D Imaging.

Area of Science:

• Biophysics
• Cell Biology
• Microscopy

Background:

• Expansion Microscopy (ExM) enables nanoscale imaging by physically expanding samples.
• Light-Sheet Fluorescence Microscopy (LSFM) provides rapid, high-resolution, volumetric imaging.
• Integrating ExM and LSFM (ExLSFM) offers synergistic benefits for biological specimen visualization.

Purpose of the Study:

• To review the combined advancements of ExM and LSFM, termed ExLSFM.
• To highlight the performance and applications of the integrated ExLSFM technique.
• To encourage wider adoption and further development of ExLSFM.

Main Methods:

• Detailed review of chemical processes critical for ExM.
• Analysis of tailored LSFM optical setups for expanded samples.
• Emphasis on sample preparation adjustments for ExLSFM data acquisition.

Main Results:

• ExLSFM demonstrates excellent performance by combining ExM and LSFM.
• The review covers diverse specimen types successfully studied with ExLSFM.
• Key chemical, optical, and sample preparation factors for ExLSFM are identified.

Conclusions:

• ExLSFM represents a powerful integrated imaging modality.
• Further development and application of ExLSFM are anticipated.
• This review aims to foster greater understanding and innovation in ExLSFM.