Abstract
This chapter introduces the principles of the fluorescent speckle microscopy method for microtubules, and then briefly describes the features of a digital imaging system that are important for speckle image recording and analysis with other optical modes, such as phase contrast or vital staining, with DNA dyes such as the blue fluorescent Hoescht or DAPI to record chromosome position. Applications of the method are presented for measuring microtubule movements in tissue cell spindles and asters and in spindles reassembled in vitro in cytoplasmic extracts of Xenopus eggs. A high-resolution, multimode, multiwavelength, digital fluorescence light microscope is used for fluorescence imaging of mitosis in living cells or cytoplasmic extracts that have enough sensitivity for fluorescence speckle imaging. Images of chromosomes and cell structure can also be obtained by transmitted light differential interference contrast (DIC) or phase contrast. The microscope utilizes conventional wide field optics and a cooled, slow-scan CCD camera for image detection.
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