Abstract
Abstract A confocal scanning Raman microscope was constructed for spectroscopy and microscopy of biological samples (Fig. 1). The microscope contains an illumination system in which a focused krypton laser beam, of which the 647.1 nm line is used to reduce damage, is scanned over the sample using a scanning mirror which moves around two orthogonal axes. Raman scattered light is collected by a water immersion objective which directs the light on the scanning mirror. Spectral analysis takes place in a monochromator with two exit ports: one for spectroscopical purposes the other one for imaging. An image is made by scanning the light, present in the Raman band passed by the monochromator, over a CCD using a second scanning mirror which moves synchronously with the first mirror. The spatial resolution is of the order of 0.3 × 0.3 × 1.2 μm3 (x,y,z). With this microscope we studied the following samples:
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