動的ホローコーン照明を用いた生物試料の位相構造観察

抄録

Phase-contrast imaging of living tissue is one of the most important techniques for investigating biological structures. We have developed phase-contrast optical microscopy by integrating dynamic hollow-cone illumination and active image processing to visualize the phase and amplitude structures of biological tissues. A figure-of-8-shaped median spatial frequency enhancement filter, which had an optical transfer function with sign correction and was split into two areas, was applied to extract separate phase and amplitude spectra of biological tissues. All spatial frequencies were covered by superimposing the figure-of-8-shaped optical transfer functions at different azimuth angles. Imaging revealed clear phase and absorptive liver structures, such as those of lymphocytes and cell nuclei. We also evaluated the imaging features of the microscope using transparent microspheres, and examined the possibility of applying it to the observation of thick biological tissue by reducing nonlinear imaging components.