Observations using conventional microscopy often lack information related to the fine structures of an object, such as intensity and phase, because of limitations in the lens aperture. In this study, the intensity and phase information are appropriately converted into an electrical signal using laser scanning and photodetectors. Intensity and phase are completely separated, and the missing information is restored based on the frequency of the electrical signal. Using this method, the original intensity and phase information of the object to be observed can be correctly restored. Therefore, we propose a novel method to calculate the degree of intensity and phase modulation by calculating the direct and alternating current components obtained from the output of the sum and difference of the two photodetectors. The degree of spatial frequency modulation is corrected according to the electrical signal frequency to detect transparent or unstained cells. We first performed laser scanning of an object. Then, signals were detected using two photodetectors placed in the far-field, separated by the optical axis as the boundary. The output signals of the photodetectors were processed and the intensity and phase were unambiguously separated, thus allowing the visualization of the phase information of the transparent bodies and unstained cells. Spatial frequency correction was performed to correct the modulation. Our method successfully separated the information related to the intensity and optical path difference (OPD). In future work, by accurately correcting the intensity and OPD, it will be possible to separate the absorption rate from the ratio of the irradiation light intensity to the observed intensity and to separate the OPD into the refractive index and the thickness information. This method allows the accurate determination of these parameters in a noninvasive manner.

中文翻译：

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扫描激光显微镜中MTF的校正和量化

由于镜头孔径的限制，使用传统显微镜进行的观察通常缺乏与物体精细结构相关的信息，例如强度和相位。在这项研究中，使用激光扫描和光电探测器将强度和相位信息适当地转换为电信号。强度和相位完全分离，根据电信号的频率恢复缺失的信息。使用这种方法，可以正确还原被观察物体的原始强度和相位信息。因此，我们提出了一种新的方法，通过计算从两个光电探测器的和和差的输出中获得的直流和交流分量来计算强度和相位调制的程度。根据电信号频率校正空间频率调制的程度，以检测透明或未染色的细胞。我们首先对物体进行激光扫描。然后，使用放置在远场中的两个光电探测器检测信号，以光轴为边界。光电探测器的输出信号经过处理，强度和相位被明确分离，从而使透明体和未染色细胞的相位信息可视化。进行空间频率校正以校正调制。我们的方法成功地分离了与强度和光程差 (OPD) 相关的信息。在未来的工作中，通过准确校正强度和 OPD，可以将吸收率从照射光强度与观察强度的比率中分​​离出来，并将 OPD 分离为折射率和厚度信息。该方法允许以无创方式准确确定这些参数。