Optimised tissue clearing minimises distortion and destruction during tissue delipidation.

Area of Science:

• Biotechnology
• Histology
• Microscopy

Background:

• Tissue clearing techniques aim to render intact tissues transparent for imaging.
• Thicker samples often require delipidation before refractive index (RI)-matching, but precise control is challenging.
• Excessive delipidation can cause tissue distortion and protein loss, hindering accurate analysis.

Purpose of the Study:

• To develop a tissue clearing strategy for better monitoring and precise determination of delipidation.
• To optimize the balance between lipid removal and tissue integrity for enhanced transparency.

Main Methods:

• Developed Accurate delipidation with Optimal Clearing (Accu-OptiClearing) by combining sodium dodecyl sulphate (SDS) with the RI-matching solution OPTIClear.
• Assessed clearing rate, protein loss, tissue distortion, and antigen preservation.
• Evaluated the preview of final tissue transparency before final RI-matching immersion.

Main Results:

• Accu-OptiClearing accelerated tissue clearing and preserved tissue structure and antigens better than SDS delipidation alone.
• Accu-OptiClearing retained more lipids while achieving similar transparency compared to SDS.
• The method allowed for better preview of final transparency, indicating optimal delipidation.

Conclusions:

• Accu-OptiClearing, combining OPTIClear with detergents, avoids excessive delipidation.
• This strategy leads to accelerated tissue clearing, reduced tissue damage, and improved antigen preservation.
• Accu-OptiClearing offers a more controlled approach to tissue clearing for advanced biological imaging.