Kaushalya Sanjeev Kumar, Balaji Jayaprakash, Garai Kanchan, Maiti Sudipta
Tata Institute of Fundamental Research, Homi Bhabha Road, Colaba, Mumbai 400005, India.
Appl Opt. 2005 Jun 1;44(16):3262-5. doi: 10.1364/ao.44.003262.
In confocal fluorescence correlation microscopy (FCM) it is important to ensure that the correlation measurement is actually performed at the chosen location of the three-dimensional image of the specimen. We present a confocal FCM design that provides an automatic real-time readout of the location in the confocal microscopic image, which is aligned with the detector of the fluorescence correlation spectrometer. The design accomplishes this without using any special positioning device. The design is based on an apertured fluorescence detector placed close to the back aperture of the objective lens and can be easily incorporated into virtually any confocal microscope. We demonstrate the method by performing FCM measurements of a dye diffusing on a cell membrane.
在共聚焦荧光相关显微镜（FCM）中，确保相关测量实际上是在标本三维图像的选定位置进行非常重要。我们提出了一种共聚焦FCM设计，该设计可提供共聚焦显微镜图像中位置的自动实时读数，该位置与荧光相关光谱仪的探测器对齐。该设计无需使用任何特殊定位装置即可实现这一点。该设计基于一个靠近物镜后孔径放置的带孔荧光探测器，并且可以很容易地集成到几乎任何共聚焦显微镜中。我们通过对细胞膜上扩散的染料进行FCM测量来演示该方法。
