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Arrangement for confocal fluorescence microscopy

  • US 5,751,417 A
  • Filed: 03/20/1996
  • Issued: 05/12/1998
  • Est. Priority Date: 03/20/1995
  • Status: Expired due to Term
First Claim
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1. An arrangement for confocal and quasifocal fluorescence microscopy in which the following are arranged in succession in an image beam path:

  • (a) an objective arrangement for acquiring an image of a specimen to be examined;

    (b) at least one scanner mirror located below said objective arrangement;

    (c) a tube lens for producing an image of said specimen in an image plane;

    (d) a first confocal diaphragm located in said image plane of said tube lens;

    (e) a first spectrometer arrangement with(i) a first lens for imaging an opening of said diaphragm at infinity,(ii) a dispersion prism for spectral decomposition of light emerging from said first lens, and(iii) a second lens for producing an image in an exit slit plane of said first spectrometer arrangement which represents an intermediate image plane;

    (f) a wavelength selection diaphragm located in said exit slit plane with a host of parallel slits with a slit width or spacing which corresponds to a desired bandwidth and which can be shifted perpendicularly to a longitudinal direction of the slits for purposes of selection of an emission wavelength;

    (g) a second spectrometer arrangement which is analogous to said first spectrometer arrangement and which forms a subtractive dispersion means to spectrally re-combine the light of said wavelength selection diaphragm which has been spectrally fanned out and to image it in a second image plane; and

    (h) a detector for acquiring a brightness distribution in said second image plane;

    wherein said wavelength selection diaphragm constitutes a means for coupling of excitation light by an excitation beam path travelling back along an arrangement comprised of said first spectrometer arrangement, said first confocal diaphragm, said tube lens, said scanner mirror and said objective arrangement;

    wherein the excitation beam path leads from a source of monochromatic excitation light by way of a second confocal diaphragm, which corresponds to said first confocal diaphragm, and a third spectrometer arrangement, analogous to said first spectrometer arrangement, to said wavelength selection diaphragm;

    wherein the image beam path and the excitation beam path are constituted and matched to one another in a manner causing the light emitted from said specimen to reach said detector by way of said wavelength selection diaphragm, and conversely, preventing excitation light from said wavelength selection diaphragm from hitting said detector and causing the excitation light to traverse said first confocal diaphragm and reach said specimen.

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