ACT-PRESTO: Biological tissue clearing and immunolabeling methods for volume imaging

Eunsoo Lee, Woong Sun

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)


The identification and exploration of the detailed organization of organs or of the whole body at the cellular level are fundamental challenges in biology. Transitional methods require a substantial amount of time and effort to obtain a 3D image and including sectioning the intact tissue, immunolabeling, and imaging serially-sectioned tissue, which produces a loss of information at each step of the process. In recently developed approaches for high-resolution imaging within intact tissue, molecular characterization has been restricted to the labeling of proteins. However, currently available protocols for organ clearing require a considerably long process time, making it difficult to implement tissue clearing techniques in the lab. We recently established a rapid and highly-reproducible protocol termed ACT-PRESTO (active clarity technique–pressure related efficient and stable transfer of macromolecules into organs), which allows for tissue clearance within several hours. Moreover, ACT-PRESTO enables rapid immunolabeling with conventional methods and accelerates antibody penetration into the deep layer of densely-formed, thick specimens by applying pressure or convection flow. We describe how to prepare tissues, how to clear by lipid removal using electrophoresis, and how to immuno-stain by a pressure-assisted delivery. The rapidity and consistency of the protocol will expedite the performance of 3D histological research and volume-based diagnoses.

Original languageEnglish
Article numbere54904
JournalJournal of Visualized Experiments
Issue number118
Publication statusPublished - 2016 Dec 31

Bibliographical note

Funding Information:
This research was supported by the Brain Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Science, ICT, and Future Planning (NRF-2015M3C7A1028790).

Publisher Copyright:
© 2016 Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License.


  • 3D imaging
  • Bioengineering
  • Immunolabeling
  • Issue 118
  • Neuroscience
  • Tissue clearing
  • Tissue engineering
  • Tissue-hydrogel

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry,Genetics and Molecular Biology
  • General Immunology and Microbiology


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