A MODIFIED DIFFERENTIAL STAIN FOR CARTILAGE AND BONE IN WHOLE MOUNT PREPARATIONS OF MAMMALIAN FETUSES AND SMALL VERTEBRATES
A MODIFIED DIFFERENTIAL STAIN FOR CARTILAGE AND BONE IN WHOLE MOUNT PREPARATIONS OF MAMMALIAN FETUSES AND SMALL VERTEBRATES
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DOI：
10.3109/10520298309066773
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发表时间：
1983-01-01
期刊：
影响因子：
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通讯作者：
BRYDEN, MM
中科院分区：
文献类型：
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作者：
KELLY, WL;BRYDEN, MM
用福尔马林-乙酸乙酯-乙醇固定，避免了阿尔新蓝和茜素红对脊椎动物整体骨骼的染色，极大地促进了该方法的酶清除步骤。这种改进的方法在胚胎和新生小鼠身上进行了测试，并在包括硬骨鱼、爬行动物、两栖动物和鸟类在内的各种脊椎动物上进行了测试，结果表明，该方法快速、可重复性和永久性。这种方法不是那么快，但至少在某些情况下是优越的。在福尔马林-乙酸乙酯-乙醇中固定40min，阿利新蓝8GX染色，然后用胰酶透明，可获得最佳结果。37℃孵化显著缩短了完成后一步所需的时间。C.下一步是用茜素红S在KOH弱溶液中对骨骼进行染色，然后在KOH-甘油系列中清除。
Fixation in formol-acetic-alcohol as a preclude to the staining of whole mount vertebrate skeletons with alcian blue and alizarin red S has greatly facilitated the enzyme clearing step of the method. The modified method was tested on fetal and neonatal mice, and on a variety of vertebrates including bony fish, reptiles, amphibia and birds, and shown to be rapid, reproducible and permanent. The method is not so rapid but is superior at least in certain circumstances. Optimal results were obtained by fixing in formol-acetic-alcohol for 40 min, staining cartilage with alcian blue 8GX, then clearing with trypsin. The time taken to complete the latter step was reduced significantly by incubation at 37.degree. C. The next step was to stain bone using alizarin red S in a weak solution of KOH, followed by clearing in a KOH-glycerol series.