Schulich Faculty of Chemistry Technion, Israel Institute of Technology, Haifa, Israel.
Biophys J. 2010 Aug 4;99(3):971-8. doi: 10.1016/j.bpj.2010.05.002.
We present what is, to our knowledge, a new methodology for high-resolution three-dimensional imaging of oxygen concentration near live cells. The cells are placed in the buffer solution of a stable paramagnetic probe, and electron spin-resonance microimaging is employed to map out the probe's spin-spin relaxation time (T(2)). This information is directly linked to the concentration of the oxygen molecule. The method is demonstrated with a test sample and with a small amount of live photosynthetic cells (cyanobacteria), under conditions of darkness and light. Spatial resolution of approximately 30 x 30 x 100 microm is demonstrated, with approximately microM oxygen concentration sensitivity and sub-fmol absolute oxygen sensitivity per voxel. The use of electron spin-resonance microimaging for oxygen mapping near cells complements the currently available techniques based on microelectrodes or fluorescence/phosphorescence. Furthermore, with the proper paramagnetic probe, it will also be readily applicable for intracellular oxygen microimaging, a capability which other methods find very difficult to achieve.
我们提出了一种新的方法，用于对活细胞附近的氧浓度进行高分辨率三维成像。将细胞置于稳定顺磁探针的缓冲溶液中，并采用电子自旋共振微成像技术来绘制探针的自旋-自旋弛豫时间（T(2)）。该信息与氧分子的浓度直接相关。该方法通过测试样本和少量活体光合细胞（蓝藻）进行了演示，分别在黑暗和光照条件下进行。证明了大约 30 x 30 x 100 微米的空间分辨率，具有大约 microM 氧浓度灵敏度和每个体素的亚飞摩尔绝对氧灵敏度。电子自旋共振微成像在细胞附近进行氧测绘的应用补充了目前基于微电极或荧光/磷光的可用技术。此外，使用适当的顺磁探针，它也将很容易适用于细胞内氧微成像，而其他方法很难实现这一功能。
