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Microscope alignment using real-time Imaging FCS
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DOI：10.1016/j.bpj.2022.06.009.png)
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Abstract
En 中文
Modern electron-multiplying charge-coupled device (EMCCD) and scientific complementary metal-oxide semi-conductor (sCMOS) cameras read out fluorescence data with single-molecule sensitivity at thousands of frames per second. Exploiting these capabilities in full requires data evaluation in real time. The direct camera-read-out tool presented here allows access to the data while the camera is recording. This provides simplified and accurate alignment procedures for total internal reflection fluorescence microscopy (TIRFM) and single-plane illumination microscopy (SPIM), and simplifies and accelerates fluorescence experiments. The tool handles a range of widely used EMCCD and sCMOS cameras and uses imaging fluores-cence correlation spectroscopy for its evaluation. It is easily extendable to other camera models and other techniques and is a base for automated TIRFM and SPIM data acquisition.
Keywords:
CORRELATION SPECTROSCOPY
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