Confocal laser microscope (CLM) is indispensable today in the biology research field as the tool to clarify three
dimensional structure and temporal transformations of living cells. The biggest advantage of CLM is to obtain
"Optical slice images" in the direction of depth. The fluorescence from specimen is detected by a photo-detector in
CLM through the small aperture called "Pinhole". The smaller the diameter of the pinhole is, the thinner the optical
slice becomes. However, there is a problem that the contrast degrades because the images darken as the pinhole
gets smaller, while the out-of-focus light increases as the pinhole is enlarged.
To solve the problem, we developed a new detection method. In this method named "VAAS", it provides with the
detector that captures light that doesn't pass through the pinhole in addition to the detector that captures light passes
through the pinhole. Both detectors convert the light into electric signals at the same time. This method enables to
eliminate out-of-focus light from the bright images acquired with large pinhole. In addition, quantitative
experiments and analysis has proved that the contrast would be improved about 10dB compared with conventional
CLM. VAAS is expected to be applied widely in the field of research to observe living cells where the reduction of
optical toxicity is required in the future.
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