Raman mapping, with a spatial resolution close to the diffraction limit, can help to reveal the distribution of chemical species at the surface of an heterogeneous sample.As early as 1975,three methods of sample laser illumination and detector configuration have been proposed to perform Raman mapping at the microscopic level (Fig. 1),:- Point illumination:The basic design of the instrument is a classical Raman microprobe equipped with a PM tube or either a linear photodiode array or a two-dimensional CCD detector. A laser beam is focused on a very small area ,close to the diffraction limit.In order to explore the whole surface of the sample,the specimen is moved sequentially beneath the microscope by means of a motorized XY stage. For each point analyzed, a complete spectrum is obtained from which spectral information of interest is extracted for Raman image reconstruction.- Line illuminationA narrow laser line is focused onto the sample either by a cylindrical lens or by a scanning device and is optically conjugated with the entrance slit of the stigmatic spectrograph.
Raman image-activated cell sorting
