Effect of scanning rate on the image contrast in confocal microscopy for biological application

Abstract

In this research, the method how to estimate the image quality for different scanning rate is suggested and experimentally shown with the laboratory-built confocal laser scanning microscope. The confocal microscope is designed for in vivo reflectance imaging of a biological tissue, which uses the refractive index mismatch at the boundaries of a tissue to generate an image without any additional staining process. The two-dimensional scanning mechanism is built up with a polygonal mirror and a galvanometric mirror that can be controlled to operate at a specific speed. To examine the effect of scanning rate on the image contrast, confocal scanning images of a biological specimen are acquired with various scanning rate while the other conditions are kept same. The contrast of confocal microscopic image is transformed into the numeric expression to describe the relation between image contrast and scanning rate quantitatively. Results suggest some useful methodology of how to determine the allowable maximum scanning rate for the specific application of confocal microscopy.