ACT-PRESTO: Biological Tissue Clearing and Immunolabeling Methods for Volume Imaging

Area of Science:

• Biology
• Histology
• Biotechnology

Background:

• Understanding cellular organization in organs and whole bodies is crucial in biology.
• Traditional 3D imaging methods are time-consuming, involving tissue sectioning and serial imaging, leading to information loss.
• Current high-resolution intact tissue imaging techniques are limited by long processing times and protein-only molecular characterization.

Purpose of the Study:

• To introduce a rapid and reproducible protocol for tissue clearing and 3D imaging.
• To overcome the limitations of existing time-consuming organ clearing methods.
• To enable efficient molecular characterization of thick biological specimens.

Main Methods:

• Developed ACT-PRESTO (active clarity technique-pressure related efficient and stable transfer of macromolecules into organs), a novel protocol for rapid tissue clearing.
• Utilized electrophoresis for lipid removal during the clearing process.
• Employed pressure-assisted delivery for accelerated antibody penetration and immunolabeling in thick specimens.

Main Results:

• Achieved complete tissue clearance within several hours, a significant reduction compared to conventional methods.
• Enabled rapid immunolabeling using standard techniques.
• Demonstrated efficient antibody penetration into deep tissue layers of dense specimens.

Conclusions:

• ACT-PRESTO offers a fast, reproducible, and efficient solution for 3D histological research.
• The protocol facilitates volume-based diagnoses by enabling high-resolution imaging of cleared tissues.
• This technique accelerates the exploration of cellular organization in complex biological structures.