FxClear, a free-hydrogel electrophoretic tissue clearing method for rapid de-lipidation of tissues with high preservation of immunoreactivity

Jungyoon Choi, Eunsoo Lee, June Hoan Kim, Woong Sun

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)


Over the last two decades, several tissue clearing methodologies have been established that render tissues optically transparent and allow imaging of unsectioned tissues of significant volumes, thus improving the capacity to study the relationships between cell and 3D tissue architecture. Despite these technical advances, the important unsolved challenges that these methods face include complexity, time, consistency of tissue size before and after clearing, and ability to immunolabel various antibodies in cleared tissue. Here, we established very simple and fast tissue clearing protocol, FxClear, which involves acrylamide-free electrophoretic tissue clearing (ETC). By removal of the acrylamide infusion step, we were able to achieve fast reaction time, smaller tissue expansion, and higher immunoreactivity. Especially, immunoreactivity and fluorescence intensity were increased in FxClear-processed tissues compared to un-cleared tissues. Our protocol may be suitable for small-sized biopsy samples for 3D pathological examinations.

Original languageEnglish
Pages (from-to)436-445
Number of pages10
JournalExperimental Neurobiology
Issue number3
Publication statusPublished - 2019 Jun 1

Bibliographical note

Funding Information:
We greatly appreciate Drs. Sung Jin Jeong, Seung-Hae Kwon, Ms. Yu Jin Jang and Ms. Joo Yeong Jeon for helping with imaging processing and analysis. The imaging centers in the Korea Brain Research Institute (KBRI) and Korea Basic Science Institute (KBSI) supported data acquisition and image processing. This research was supported by the Brain Research Program through the National Research Foundation (NRF) funded by the Korean Ministry of Science, ICT & Future Planning (NRF-2015M3C7A1028790, 2018R1D1A1A02086190), KBRI basic research program through Korea Brain Research Institute funded by Ministry of Science and ICT (18-BR-03-01).

Publisher Copyright:
Copyright © Experimental Neurobiology 2019.


  • Brain tissue
  • Immunohistochemistry
  • Three dimensional imaging
  • Tissue engineering

ASJC Scopus subject areas

  • Clinical Neurology
  • Cellular and Molecular Neuroscience


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