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Scanning confocal microscope using fluorescence detection

  • US 5,296,703 A
  • Filed: 04/01/1992
  • Issued: 03/22/1994
  • Est. Priority Date: 04/01/1992
  • Status: Expired due to Term
First Claim
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1. A confocal microscope for scanning a sample, comprising:

  • a laser for generating a collimated incident beam having a predetermined wavelength;

    optical path means includinga primary dichroic mirror, anda first scanning mirror, disposed between the dichroic mirror and the sample and rotatable about a first axis, for scanning the incident beam repeatedly across the sample, whereupon the sample fluoresces and emits radiation having a first predetermined wavelength, different from the wavelength of the incident beam, a portion of the emitted radiation forming a fluorescence beam that retraces the path of the incident beam back to the primary dichroic mirror,wherein the primary dichroic mirror separates the fluorescence beam from the incident beam;

    a fluorescence beam detector for detecting the intensity of the fluorescence beam and for generating a corresponding fluorescence scan signal; and

    reference beam means includingmeans for directing a reference beam toward the first scanning mirror, where it impinges on the first scanning mirror at a location coincident with the incident beam, to produce a scanning reference beam,a grating having a series of alternating transparent and opaque regions and positioned to receive the scanning reference beam and produce a pulsed reference beam indicative of the scan rate, anda reference beam detector for detecting the intensity of the pulsed reference beam and for generating a corresponding clock signal representative of the scan rate of the scanning reference beam and thereby the incident beam;

    wherein the clock signal generated by the reference beam detector can be used to sample the fluorescence scan signal generated by the fluorescence beam detector.

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