Fast free of acrylamide clearing tissue (FACT) for clearing, immunolabelling and three-dimensional imaging of partridge tissues

Autor: Ali Niazi, Farhad Rahmanifar, Farzad Mohammad Rezazadeh, Amin Tamadon, Kazuyoshi Tsutsui, Saman Saedi, Mohammad Reza Namavar, Mohammad Reza Jafarzadeh Shirazi, Ali Arabi Monfared, Mehdi Dianatpour, Amir Akhlaghi, Nader Tanideh
Rok vydání: 2018
Předmět:
Zdroj: Microscopy Research and Technique. 81:1374-1382
ISSN: 1059-910X
DOI: 10.1002/jemt.23078
Popis: Fast free of acrylamide clearing tissue (FACT) is a modified sodium dodecyl sulfate-based clearing protocol for the chemical clearing of lipids that completely preserves fluorescent signals in the cleared tissues. The FACT protocol was optimized to image translucent immunostained brain and non-nervous tissues. For this purpose adult male Chukar partridge (Alectoris chukar) was used as a model. After clearing the tissues, 1 or 2 mm-thickness sections of tissues were immunolabeled. The paraventricular nucleus in the hypothalamus (2-mm section) was cleared with FACT, and then was stained with gonadotropin-inhibitory hormone (GnIH) antibody and Hoechst. Simultaneously, immunohistochemical (IHC) staining of cryosectioned brain (30 μm) was done by GnIH-antibody. The FACT protocol and staining of cell nuclei of nine other tissues were done by a z-stack motorized fluorescent microscope. GnIH-immunoreactive neurons were found by FACT and IHC during the breeding season in male partridges. Deep imaging of the kidney, duodenum, jejunum, lung, pancreas, esophagus, skeletal muscle, trachea, and testis were also done. The FACT protocol can be used for the three-dimensional imaging of various tissues and immunostained evaluation of protein markers. RESEARCH HIGHLIGHT: The FACT is a simple and cheap method for whole tissue clearing. The FACT-cleared tissues can be imaged with simple fluorescent microscopes. For the first time, using the FACT, three-dimensional imaging of various tissues was done.
Databáze: OpenAIRE
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