Université de Bordeaux 2, CNRS UMR 5084, B8 Avenue des Facultés, 33405 Pessac-Cedex, France.
Anal Bioanal Chem. 2010 Jul;397(6):2123-9. doi: 10.1007/s00216-010-3817-2. Epub 2010 Jun 2.
FTIR microscopy with a focal plane array (FPA) of detectors enables routine chemical imaging on individual cells in only a few minutes. The brilliance of synchrotron radiation (SR) IR sources may enhance the signal obtained from such small biosamples containing small amounts of organic matter. We investigated individual cells obtained from a cell culture specifically developed for transmission FTIR imaging using either a Globar or an SR source coupled to the same instrumentation. SR-IR source focussing was optimized to control the energy distribution on the FPA of detectors. Here we show that accessing the IR absorption distribution from all the organic contents of cells at 1 x 1 microm pixel resolution was possible only with high circulating current (> or = 1.2 A) illuminating a limited number of the FPA's detectors to increase the signal-to-noise ratio of IR images. Finally, a high-current SR ring is mandatory for collecting FTIR images of biosamples with a high contrast in minutes.
傅里叶变换红外显微镜与焦平面阵列(FPA)探测器结合使用,仅需几分钟即可对单个细胞进行常规化学成像。同步辐射(SR)红外源的亮度可能会增强从含有少量有机物的此类小生物样本中获得的信号。我们使用 Globar 或 SR 源研究了专门为传输傅里叶变换红外成像开发的细胞培养物中获得的单个细胞,这些源都连接到相同的仪器上。优化了 SR-IR 源聚焦,以控制探测器 FPA 上的能量分布。在这里,我们表明仅当以 > 或 = 1.2 A 的高循环电流照射有限数量的 FPA 探测器以增加红外图像的信噪比时,才能以 1 x 1 微米像素分辨率从细胞的所有有机物质中获取红外吸收分布。最后,对于在几分钟内收集具有高对比度的生物样本的傅里叶变换红外图像,高电流 SR 环是必需的。