ACT-PRESTO

Biological tissue clearing and immunolabeling methods for volume imaging

Eunsoo Lee, Woong Sun

Research output: Contribution to journalArticle

4 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

Fingerprint

Biological organs
Macromolecules
Tissue
Imaging techniques
Pressure
Convection
Electrophoresis
Antibodies
Labeling
Lipids
Coloring Agents
Proteins

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

Cite this

ACT-PRESTO : Biological tissue clearing and immunolabeling methods for volume imaging. / Lee, Eunsoo; Sun, Woong.

In: Journal of Visualized Experiments, Vol. 2016, No. 118, e54904, 31.12.2016.

Research output: Contribution to journalArticle

@article{165693fc273e4660a59857c0be197c27,
title = "ACT-PRESTO: Biological tissue clearing and immunolabeling methods for volume imaging",
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.",
keywords = "3D imaging, ACT-PRESTO, Bioengineering, Immunolabeling, Issue 118, Neuroscience, Tissue clearing, Tissue engineering, Tissue-hydrogel, X-CLARITY",
author = "Eunsoo Lee and Woong Sun",
year = "2016",
month = "12",
day = "31",
doi = "10.3791/54904",
language = "English",
volume = "2016",
journal = "Journal of Visualized Experiments",
issn = "1940-087X",
publisher = "MYJoVE Corporation",
number = "118",

}

TY - JOUR

T1 - ACT-PRESTO

T2 - Biological tissue clearing and immunolabeling methods for volume imaging

AU - Lee, Eunsoo

AU - Sun, Woong

PY - 2016/12/31

Y1 - 2016/12/31

N2 - 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.

AB - 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.

KW - 3D imaging

KW - ACT-PRESTO

KW - Bioengineering

KW - Immunolabeling

KW - Issue 118

KW - Neuroscience

KW - Tissue clearing

KW - Tissue engineering

KW - Tissue-hydrogel

KW - X-CLARITY

UR - http://www.scopus.com/inward/record.url?scp=85008686905&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85008686905&partnerID=8YFLogxK

U2 - 10.3791/54904

DO - 10.3791/54904

M3 - Article

VL - 2016

JO - Journal of Visualized Experiments

JF - Journal of Visualized Experiments

SN - 1940-087X

IS - 118

M1 - e54904

ER -