Simultaneous measurement of Ca2+ and cellular dynamics:: Combined scanning ion conductance and optical microscopy to study contracting cardiac myocytes
Simultaneous measurement of Ca2+ and cellular dynamics:: Combined scanning ion conductance and optical microscopy to study contracting cardiac myocytes
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DOI：
10.1016/s0006-3495(01)75826-2
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发表时间：
2001-09-01
影响因子：
3.4
通讯作者：
Korchev, YE
中科院分区：
文献类型：
--
作者：
Shevchuk, AI;Gorelik, J;Korchev, YE
我们开发了一种用于扫描离子电导显微镜的距离调制协议，为成像收缩细胞提供了一个健壮而可靠的距离控制机制。这项技术可以测量细胞高度从10纳米到几微米的快速变化，时间分辨率为毫秒级。这一点已经在心肌细胞收缩的极端情况下得到了证明。通过将这种方法与激光共聚焦显微镜相结合，可以同时测量心肌细胞的纳米运动和细胞膜下的局部钙浓度。尽管细胞运动很大，但同时跟踪细胞高度的变化和测量细胞表面附近的细胞内钙离子是可能的，同时保持细胞的功能。
We have developed a distance modulated protocol for scanning ion conductance microscopy to provide a robust and reliable distance control mechanism for imaging contracting cells. The technique can measure rapid changes in cell height from 10 nm to several micrometers, with millisecond time resolution. This has been demonstrated on the extreme case of a contracting cardiac myocyte. By combining this method with laser confocal microscopy, it was possible to simultaneously measure the nanometric motion of the cardiac myocyte, and the local calcium concentration just under the cell membrane. Despite large cellular movement, simultaneous tracking of the changes in cell height and measurement of the intracellular Ca2+ near the cell surface is possible while retaining the cell functionality.