Use of a fluorescent cyanine dye for electrophysiological studies on the frog cornea.
Use of a fluorescent cyanine dye for electrophysiological studies on the frog cornea.
复制标题
使用荧光花青染料对青蛙角膜进行电生理学研究。
DOI：
10.1152/ajpcell.1980.238.1.c21
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发表时间：
1980
期刊：
影响因子：
--
通讯作者：
W. S. Rehm
中科院分区：
文献类型：
--
作者：
C. Graves;G. Sachs;W. S. Rehm
材料与方法本研究使用牛蛙（Rana catesbeiana）角膜。将断头青蛙的头部置于含有青蛙林格氏溶液的培养皿中。通过在角膜-巩膜接合处后约1-2 mm处围绕地球仪切开一个切口，沿着虹膜和透镜去除角膜。从角膜中取出透镜和虹膜。将灌注室（图1B）置于林格溶液中，并将角膜浮在灌注室的轮廓表面上。然后将腔室从林格氏溶液中取出，并在腔室上的凹槽处用细缝合线牢固地系紧角膜。将黑色树脂玻璃环放置在角膜边缘上的腔室肩部上，并将矿物油和凡士林（约15）的粘性混合物施加到环和角膜之间的区域。这在角膜的水侧和泪侧之间提供了电密封。在角膜的水侧保持约20 cmH&I的流体静压。然后将体外室安装在泪液储存器中（图1A），将其置于荧光分光光度计的样品室中。角膜的水侧以2 ml/min的速率用由Buchler peristartic泵递送的林格溶液灌注。将该室浸没在储液器中，将含有4 μ m花青染料diS-Ca-（5）的林格溶液注入到水性室中约10
MATERIALS AND METHODSThe corneas of bullfrogs (Rana catesbeiana) were used for this study. The head of a decapitated frog was placed in a petri dish containing frog Ringer solution. The cornea, along with the iris and lens, was removed by making an incision around the globe about 1-2 mm posterior to the corneal-scleral junction. The lens and iris were removed from the cornea. A perfusion chamber (Fig. 1B) was placed in Ringer solution and the cornea was floated onto the contoured surface of the chamber. The chamber was then removed from the Ringer solution, and the cornea was tied securely with fine suture thread at the groove on the chamber. A black Plexiglas ring was placed over the edges of the cornea on to the shoulder of the chamber and a viscous mixture of mineral oil and Vaseline (about 15) was applied to the area between the ring and the cornea. This provided an electrical seal between the aqueous side and tear side of the cornea. A hydrostatic pressure of approximately 20 cmH&I was maintained on the aqueous side of the cornea, The in vitro chamber was then mounted in a tear fluid reservoir (Fig. IA), which was placed into the sample compartment of a spectrofluorometer. The aqueous side of the cornea was perfused at a rate of 2 ml/min with Ringer solution delivered by a Buchler peristartic pump. The chamber was immersed in a reservoir and a Ringer solution containing 4 pm of cyanine dye, diS-Ca-(5), was infused into the aqueous chamber for about 10