Analysis of point source size on measurement accuracy of lateral point-spread function of confocal Raman microscopy

Abstract

Confocal Raman Microscopy (CRM) has matured to become one of the most powerful instruments in analytical science because of its molecular sensitivity and high spatial resolution. Compared with conventional Raman Microscopy, CRM can perform three dimensions mapping of tiny samples and has the advantage of high spatial resolution thanking to the unique pinhole. With the wide application of the instrument, there is a growing requirement for the evaluation of the imaging performance of the system. Point-spread function (PSF) is an important approach to the evaluation of imaging capability of an optical instrument. Among a variety of measurement methods of PSF, the point source method has been widely used because it is easy to operate and the measurement results are approximate to the true PSF. In the point source method, the point source size has a significant impact on the final measurement accuracy. In this paper, the influence of the point source sizes on the measurement accuracy of PSF is analyzed and verified experimentally. A theoretical model of the lateral PSF for CRM is established and the effect of point source size on full-width at half maximum of lateral PSF is simulated. For long-term preservation and measurement convenience, PSF measurement phantom using polydimethylsiloxane resin, doped with different sizes of polystyrene microspheres is designed. The PSF of CRM with different sizes of microspheres are measured and the results are compared with the simulation results. The results provide a guide for measuring the PSF of the CRM.