An Approach to Measuring the Point Spread Function of the Confocal Raman Microscope

The confocal Raman microscope (CRM) is a powerful tool in analytical science. Image quality is the most important performance indicator of CRM systems. The point spread function (PSF) is one of the most useful tools to evaluate the image quality of microscopic systems. A method based on a point-like object is proposed to measure the PSF of CRM, and the size effect of spherical objects is discussed. A series of phantoms are fabricated by embedding different sizes of polystyrene microspheres into polydimethylsiloxane matrix. The diameters of microspheres are from 0.2 µm to 5 µm. The phantoms are tested by measuring the PSF of a commercial CRM whose nominal lateral resolution is about 1 µm. Results of the PSF are obtained and the accuracy of resolution is used to evaluate the size effect of the microspheres. Experimental results are well consistent with theoretical analysis. The error of the PSF can be decreased by reducing the diameter of the microsphere but meanwhile the signal-to-noise ratio (S/N) will be lowered as well. The proper diameter of microspheres is proposed in consideration of the trade-off between the S/N and the measurement error of the PSF. Results indicate that the method provides a useful approach to measurement of the PSF and the resolution of the CRM.