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

Eunsoo Lee, Woong Sun

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

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
Volume2016
Issue number118
DOIs
Publication statusPublished - 2016 Dec 31

Keywords

  • 3D imaging
  • ACT-PRESTO
  • Bioengineering
  • Immunolabeling
  • Issue 118
  • Neuroscience
  • Tissue clearing
  • Tissue engineering
  • Tissue-hydrogel
  • X-CLARITY

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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