Mohammad Rezazadeh, Farzad and Saedi, Saman and Rahmanifar, Farhad and Namavar, Mohammad Reza and Dianatpour, Mehdi and Tanideh, Nader and Akhlaghi, Amir and Niazi, Ali and Arabi Monfared8, Ali and Tsutsui, Kazuyoshi and Jafarzadeh Shirazi, Mohammad Reza and Tamadon, Amin (2018) Fast free of acrylamide clearing tissue (FACT) for clearing, immunolabelling and three-dimensional imaging of partridge tissues. Microsc Res Tech..
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Abstract
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.
| Item Type: | Article |
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| Subjects: | R Medicine > R Medicine (General) |
| Divisions: | Faculty of Medicin |
| Depositing User: | زهرا صفایی |
| Date Deposited: | 17 Apr 2019 08:13 |
| Last Modified: | 17 Apr 2019 08:13 |
| URI: | http://eprints.bpums.ac.ir/id/eprint/7753 |
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