Abstract
In merits of super-resolved resolution and fast speed of three-dimensional (3D) optical sectioning capability, structured illumination microscopy (SIM) has found variety of applications in biomedical imaging. So far, most SIM systems use monochrome CCD or CMOS cameras to acquire images and discard the natural color information of the specimens. Although multicolor integration scheme are employed, multiple excitation sources and detectors are required and the spectral information is limited to a few of wavelengths. Here, we report a new method for full-color SIM with a color digital camera. A data processing algorithm based on HSV (Hue, Saturation, and Value) color space is proposed, in which the recorded color raw images are processed in the Hue, Saturation, Value color channels, and then reconstructed to a 3D image with full color. We demonstrated some 3D optical sectioning results on samples such as mixed pollen grains, insects, micro-chips and the surface of coins. The presented technique is applicable to some circumstance where color information plays crucial roles, such as in materials science and surface morphology.
Highlights
The color digital camera can produce color image is due to the use of Bayer filter
For the multicolor microscopes based on the multi-channel integration geometry, there is a narrow band-pass filter put in front of each detector, the integrated image cannot restore the full-color information of the specimen
To solve the above issue for a full-color structured illumination optical sectioning microscopy, we propose a new scheme based on our developed Digital Micro-mirror Device (DMD)-based LED-illumination SIM system[16], in which a color CMOS camera and a white-light LED illumination is employed to realize SIM with full natural color
Summary
In order to evaluate the spatial resolution of the color SIM microscope, we used 170 nm in diameter green fluorescent microspheres (520 nm emission@475 nm excitation) as test samples. Nguyen et al.[29] proposed a method based on multi-view image fusion scheme for capture of natural-color 3D models of insects It used the color texture extracted from the specimen to map the 3D model of the reconstructed object. We have proposed a scheme for full-color structured illumination microscopy (C-SIM) based on DMD fringe projection and LED illumination, which has practical value to acquiring the broadband fluorescence signals and natural color of reflected light from object surfaces in three dimensions, rather than the artificial color that usually generated from gray-scale data post-processing. We have demonstrated some 3D optical sectioning results on samples such as mixed pollen grains, insects, micro-chips and metallic surface of coins This technique may find potential applications in such fields as biology (e.g. the study of structural color mechanism of animal31), materials science, microelectronics, where color information may play crucial roles
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