Tailored Sample Mounting for Light-Sheet Fluorescence Microscopy of Clarified Specimens by Polydimethylsiloxane...

Area of Science:

• Neuroscience
• Biomedical Engineering
• Microscopy

Background:

• Light-sheet fluorescence microscopy (LSFM) enables whole-organ scale imaging with microscopic resolution.
• LSFM preserves organ architecture, crucial for studying long-range neuronal circuits.
• Current sample mounting methods for LSFM, including quartz cuvettes, can be expensive and may cause damage.

Purpose of the Study:

• To develop and validate a cost-effective sample mounting solution for LSFM.
• To introduce custom-made polydimethylsiloxane (PDMS) cuvettes as an alternative to commercial quartz cuvettes.
• To evaluate the performance of PDMS cuvettes in terms of cost, image quality, and sample integrity.

Main Methods:

• Fabrication of custom polydimethylsiloxane (PDMS) cuvettes.
• Measurement of light transmittance for PDMS and quartz cuvettes.
• Whole mouse brain imaging using LSFM with both PDMS and quartz cuvettes.
• Comparison of imaging results, including image quality and sample integrity.

Main Results:

• PDMS cuvettes demonstrated comparable light transmittance to quartz cuvettes.
• LSFM imaging with PDMS cuvettes yielded high-quality images of whole mouse brains.
• The use of PDMS cuvettes with an inexpensive refractive index (RI) matching solution significantly reduced overall imaging costs.
• PDMS cuvettes, combined with the RI matching solution, caused moderate sample expansion, potentially beneficial for visualizing fine neuronal processes.

Conclusions:

• Custom-made PDMS cuvettes are a cost-effective and high-performing alternative for sample mounting in LSFM.
• PDMS cuvettes offer advantages in terms of cost reduction, image quality, and sample preservation compared to existing methods.
• This approach facilitates advanced whole-organ imaging for neuroscience research.