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ACT-PRESTO: Rapid and consistent tissue clearing and labeling method for 3-dimensional (3D) imaging.
Eunsoo Lee1, Jungyoon Choi1, Youhwa Jo1
1Department of Anatomy and Division of Brain Korea 21 Plus Program for Biomedical Science, Korea University College of Medicine, Anam-dong, Seongbuk-gu, Seoul 136-705, Korea.
Scientific Reports
|January 12, 2016
Summary
We developed ACT-PRESTO, a rapid tissue clearing method for whole organs and bodies. This technique enables fast, reproducible 3D visualization of cellular structures, preserving proteins and improving antibody penetration.
Area of Science:
- Cellular Biology
- Neuroscience
- Biotechnology
Background:
- 3D structural analysis of organs and organisms is crucial but challenging.
- Traditional methods like serial sectioning are labor-intensive, slow, and error-prone.
- Existing tissue clearing techniques offer whole-organ visualization but require lengthy processing times.
Purpose of the Study:
- To introduce a novel, rapid tissue clearing method.
- To overcome the time limitations of current tissue clearing protocols.
- To enable high-content mapping and analysis of intact tissues.
Main Methods:
- Developed the ACT-PRESTO (active clarity technique-pressure related efficient and stable transfer of macromolecules into organs) method.
- Utilized pressure to expedite antibody penetration into thick specimens.
- Ensured compatibility with conventional immunolabeling techniques.
Main Results:
- ACT-PRESTO clears tissues or whole bodies within 1 day.
- The method preserves tissue architecture and protein-based signals (e.g., fluorescent proteins).
- Achieved highly reproducible results with rapid processing.
Conclusions:
- ACT-PRESTO significantly reduces tissue clearing time while maintaining structural and molecular integrity.
- The method facilitates efficient immunolabeling in thick specimens.
- Enables high-content analysis of normal and pathological features in intact organs and bodies.
