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Fluorescence microscopy

  • US 4,791,310 A
  • Filed: 10/02/1986
  • Issued: 12/13/1988
  • Est. Priority Date: 10/02/1986
  • Status: Expired due to Term
First Claim
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1. A multi-chronal fluorescence imaging apparatus for spatial differentiation of a multitude of separate biological, organic and inorganic components that includesa sample holder for containing a sample with a plurality of fluorescent dyes bonded to specific sample components,said dyes each reaching a different maximum output intensity upon being excited and having substantially different decay times with the dye of greatest maximum output intensity reaching final decay first and the remaining dyes decaying in the order of their respective maximum output intensities such that the maximum output of each dye can be detected in an ordered sequence with little interference from the other dyes,a pulsed light source for periodically irradiating a sample in said holder with a pulsed excitation radiation wherein each dye emits fluorescent radiation at about its maximum output before reaching final decay.a spatially sensitive detector means positioned in an optical light path for viewing the radiation emitted by said dyes,a gate means having a shutter movable between a closed position wherein radiation emitted by said sample is prevented from reaching the detector means and an open position wherein radiation emitted by said sample is transmitted to the detector means,means for intermittently energizing the light source to produce a light pulse having a duration that is less than the shortest decay time of said dyes, andcontrol means to open the shutter at periodic intervals to permit the detector to view time resolved images produced by each dye prior to its decay.

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