Trial of whole-organ and whole-body clearing in fish
Trial of whole-organ and whole-body clearing in fish
复制标题
鱼类全器官和全身清除试验
DOI：
10.11369/jji.17-045
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
宇田川伸吾・矢田文・竹村明洋
中科院分区：
文献类型：
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作者：
Mahardini A;Takeuchi Y;Dinda R;Takekata H;Takemura A;宇田川伸吾・矢田文・竹村明洋
采用新研制的组织净化脱色剂ScaleCUBIC-1对马拉巴石斑鱼（Epinealumus malabaricus）、金线蓝子鱼（Siganus guttatus）和蓝魔鬼鱼（Chrysiptera cyanea）进行了净化脱色试验。在用来自马拉巴石斑鱼尾静脉的4%多聚甲醛（PFA）灌注固定后，解剖组织（脑、心脏、脾、肝、肠、鳃、眼和肌肉），并在4 ℃下在4%PFA中后固定24小时，然后浸入ScaleCUBIC-1中。随后，发现脑、脾、肠、鳃和肌肉（但不是皮肤或鳞片）已完全清除，心脏和肝脏部分清除。由于器官的厚度，肝脏的较深区域未被清除。清眼和脱色的失败部分是由于前者的色素沉着。在ScaleCUBIC-1中浸泡5天后，对幼年金线足棘鱼和成熟蓝宝石魔鬼鱼的组织获得了类似的结果。全身清洁和脱色也试验了金线spinefoot和蓝宝石魔鬼。去除鳞片和皮肤后，将标本浸入ScaleCUBIC-1中14天，成功地使两个物种的肌肉透明和脱色。得出的结论是，ScaleCUBIC-1适用于鱼类组织（包括全身）的清洁和脱色，尽管可能需要对不同鱼类物种的程序进行修改。
ScaleCUBIC-1, the newly-developed reagent for tissue clearing and decolorization, was trialed on the Malabar grouper (Epinephelus malabaricus), goldlined spinefoot (Siganus guttatus), and sapphire devil (Chrysiptera cyanea). After perfusion fixation with 4% paraformaldehyde (PFA) from the caudal vein of the Malabar grouper, tissues (brain, heart, spleen, liver, intestine, gill, eyes, and muscle) were dissected and post-fixed in 4% PFA at 4 C for 24 hrs, before being immersed in ScaleCUBIC-1. Subsequently, the brain, spleen, intestine, gill, and muscle (but not skin or scale) were found to have been completely cleared, and the heart and liver partially so. Due to the thickness of the organ, deeper regions of the liver were not cleared. The failure of eye-clearing and decolorzation was partially due to pigmentation of the former. Similar results were obtained for tissues of the juvenile goldlined spinefoot and mature sapphire devil, following immersion in ScaleCUBIC-1 for 5 days. Whole body cleaning and decolorzation were also trialed on the goldlined spinefoot and sapphire devil. Following removal of the scales and skin, the immersion of specimens in ScaleCUBIC-1 for 14 days resulted in successful clearing and decolorzation of the muscles of both species. It was concluded that ScaleCUBIC-1 is suitable for clearing and decolorzation of fish tissues, including whole body, although modifications in the procedure may be required for different fish species.