In a confocal microscope, the illumination and detection optics are focused on the same diffraction-limited spot in the sample, which is the only spot imaged by the detector during a confocal scan. This technique allows for high-resolution imaging in thick tissues. Confocal microscopy provides a means of rejecting the out-of-focus light from the detector such that it does not contribute blur to the images being collected. In fluorescence microscopy, any dye molecules in the field of view will be stimulated, including those in out-of-focus planes. The out-of-focus light will add blur to the image reducing the resolution.
For thicker samples, where the objective lens does not have sufficient depth of focus, light from sample planes above and below the focal plane will also be detected. In light microscopy, illuminating light is passed through the sample as uniformly as possible over the field of view.
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