Abstract

As the development of biomedical researches, microscopic tools with enhanced contrast and information from multiple physical perspectives will be critical. Multi-modal digital holographic microscopy combines epifluorescence microscopy and digital holographic microscopy, and it enables simultaneous detection from a single biological sample. This paper presents utilizing multi wavelengths of fluorescent excitation and hologram recording. This is realized by using a pair of 473 nm and 532 nm lasers as excitation light, and another pair of 632.8 nm and 830 nm as hologram formation. Using two or more kinds of excitation light would increase the number of visible colors, thus multiple marking would be possible. On the other hand, measured phase value is limited to range of [-π, π] without special phase unwrapping treatment. Using different wavelengths will solve this problem by increasing measurable range. Experiments using plant cells are demonstrated the dual-wavelength excitation and dual-wavelength phase imaging. Quality of phase image affected by diffraction and absorption of intracellular components is also discussed.

© 2016 Japan Society of Applied Physics, Optical Society of America