Quantitative phase imaging of live cells using fast Fourier phase microscopy
Quantitative phase imaging of live cells using fast Fourier phase microscopy
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DOI：
10.1364/ao.46.001836
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发表时间：
2007-04-01
期刊：
影响因子：
1.9
通讯作者：
Feld, Michael S.
中科院分区：
文献类型：
--
作者：
Lue, Niyom;Choi, Wonshik;Feld, Michael S.
利用图像场在两个空间分量中的分解，可以相互控制相移，开发了一种新的定量相显微镜。这种新仪器被称为快速傅里叶相显微镜（f-FPM），与我们之前报道的傅里叶相显微镜相比，它的采集率提高了100倍。在10帧/秒或更高的采集速率下，得到的定量相位图像具有衍射极限横向分辨率和路径长度稳定性优于2 nm的特点。这些特点使得f-FPM在广泛的时间尺度上研究活细胞的结构和动力学特别有吸引力。此外，我们展示了通过数字处理仪器提供的振幅和相位信息来检查亚细胞结构的可能性。因此，我们开发了软件，可以模拟相衬和微分干涉对比显微镜图像的数值处理FPM图像。这种方法增加了数字改变两干涉光束之间相移的灵活性。与典型的商用显微镜相比，所获得的图像似乎是由可变相位对比或差干涉对比显微镜记录的，这些显微镜提供了对亚细胞结构的增强视图。(c) 2007年美国光学学会。
Using the decomposition of an image field in two spatial components that can be controllably shifted in phase with respect to each other, a new quantitative-phase microscope has been developed. The new instrument, referred to as the fast Fourier phase microscope (f-FPM), provides a factor of 100 higher acquisition rate compared with our previously reported Fourier phase microscope. The resulting quantitative-phase images are characterized by diffraction limited transverse resolution and path-length stability better than 2 nm at acquisition rates of 10 frames/s or more. These features make the f-FPM particularly appealing for investigating the structure and dynamics of live cells over a broad range of time scales. In addition, we demonstrate the possibility of examining subcellular structures by digitally processing the amplitude and phase information provided by the instrument. Thus we developed software that can emulate phase contrast and differential interference contrast microscopy images by numerically processing FPM images. This approach adds the flexibility of digitally varying the phase shift between the two interfering beams. The images obtained appear as if they were recorded by variable phase contrast or differential interference contrast microscopes that deliver an enhanced view to the subcellular structure when compared with the typical commercial microscope. (c) 2007 Optical Society of America.