Intensity distortion in dual-axis galvo mirror scanning TPF imaging system

Abstract Two-photon fluorescence (TPF) imaging system with dual-axis galvo mirror as scanning unit is built. Based on the optical path of imaging system built, mechanism of how intensity distortion generating in dual-axis galvo mirror scanning TPF imaging system is theoretically analyzed. In experiment, TPF imaging is realized with Rhodamine B sample through detecting TPF signal generated from Rhodamine B molecule at excitation of broadband femtosecond pulses. The imaging result verifies the existence of intensity distortion with distribution agreeing well with the theoretical analyzation. To eliminate the intensity distortion in TPF imaging result, a mathematical method is proposed in which a calibration matrix is calculated, As the calibration matrix is applied to the distorted imaging result of Rhodamine B sample, intensity distortion is relieved effectively and precise imaging result is achieved.