Multimodal microscope which obtains various images for the same interest area simultaneously plays an
important role in the process of comprehensive analysis for biological information. Confocal imaging for reflection and
fluorescence emission light and two-photon imaging dealing with nonlinear optical effect are operated in the unified
platform by sharing system configuration commonly used for those imaging. Reflection light, fluorescence light and
nonlinear optical signal from the same focal point are detected through separate channels and converted to respective
images. Common optical system is especially customized to satisfy their requirements considering wavelength band of
light used for imaging. Multimodal microscope is implemented and verified through multichannel images for bioexperiments.
Confocal reflection imaging for label-free specimen, confocal fluorescence imaging and two-photon
fluorescence imaging for specifically stained target are realized and make complementary analysis. The used light
signals, continuous wave light from NUV to NIR and pulsed light, is verified through imaging results of designed
multimodal microscope.
The multi-color, or spectral fluorescence microscopy has ability to detect fluorescence spectral signals which are useful
in case of studying interactions and phenomena between biological samples. Recently, commercial devices are
combining with confocal microscope so to enhance lateral resolution and to have axial direction discernment. Also
Acousto-Optic Tunable Filter(AOTF) is used instead of dichroic mirror to divide excitation and emission signals with
mininum light efficiency. In addition, AOTF is used in spectral fluorescence microscopy have many advantages, these
are very fast switching speed and high resolution in wavelength selection. However it uses acousto-optic interactions in
birefringence material, Tellurium Dioxide(TeO2), the excitation light interacts with appropriate acoustic signal so that it
is diffracted to 1 or -1 order path. But the fluorescence signals from a sample propagate in 0 order path with small
different angle according to the polarization state. In this paper, a confocal-spectral microscopy is proposed with the new
kind of spectral detector design having wavelength scanning galvano mirror. It makes possible to detect broad
wavelength fluorescence signal by single PMT with simply rotating the galvano mirror. Also a new birefringent material,
calcite(CaCO3) is used to compensate polarization effect. The proposed spectral confocal microscopy with unique
spectrometer body has many advantages in comparison with commercial devices. In terms of detection method, it can be
easily applied to other imaging modalities. Hence this system will be adapted in many applications.
Confocal scanning microscopy (CSM) needs a scanning mechanism because only one point information of specimen can
be obtained. Therefore the speed of the confocal scanning microscopy is limited by the speed of the scanning tool. To
overcome this limitation from scanning tool we propose another scanning mechanism. We make three optical probes in
the specimen under confocal condition of each point. Three optical probes are moved by beam scanning mechanism with
shared resonant scanning mirror (RM) and galvanometer driven mirror (GM). As each optical probe scan allocated
region of the specimen, information from three points is obtained simultaneously and image acquisition time is reduced.
Therefore confocal scanning microscopy with multiple optical probes is expected to have three times faster speed of the
image acquisition than conventional one. And as another use, multiple optical probes to which different light wavelength
is applied can scan whole same region respectively. It helps to obtain better contrast image in case of specimens having
different optical characteristics for specific light wavelength. In conclusion confocal scanning microscopy with multiple
optical probes is useful technique for views of image acquisition speed and image quality.
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