High precision fluorometer for measuring enzymatic substrates in tissue

Claim 1

A high precision fluorometer for biochemical measurements comprising: A. a source of a beam of radiant energy; B. cell means for holding a sample to be measured, said sample being of a type exhibiting an initial level of fluorescence when irradiated by said radiant energy; c. means for dispensing a reagent into said sample cell means to thereby change the fluorescence level of said sample relative to said initial level; D. reference cell means; E. said sample cell means and said reference cell means having at least two adjacent faces at right angles to each other, said beam entering through one of said adjacent faces and fluorescence energy exiting through the other of said adjacent faces; F. photomultiplier means for converting fluorescent energy into electrical signals; G. a movably mounted mirror means for selectively directing said beam at said sample cell means or said reference cell means, whereby said sample or said reference, respectively, will be excited by said radiant energy to fluorescence, and simultaneously directing fluorescent energy from said sample cell means or said reference cell means, respectively, toward said photomultiplier means; H. means for converting said electrical signals to digital signals; I. means for integrating said digital signals over a predetermined time interval; J. means for comparing the signals from said sample cell with the signals from said reference cell, thereby producing the ratio of sample to reference intensity; K. print-out means; L. rotatable carrier means for a plurality of sample cells, said carrier means being a circular metal block containing channel means for circulation of water at a constant temperature to thereby control the temperature of said cells; M. means for rotating said carrier means to position individual cells for dispensing said reagents thereinto and excitation by said beam; and N. control means for selectively rotating said carrier means and directing said beams and activating said means for integrating said digital signals, comparing said sample and reference signals, and printing out the desired information.

Claim 5

A method for high precision biochemical analysis comprising: A. providing a source of a beam of radiant energy; B. providing a movable mirror means for selectively directing said beam toward a first position and a second position, respectively when irradiated by said radiant energy; C. providing a sample of a type exhibiting an initial level of fluorescence to be measured at said first position; D. providing a reference in said second position; E. providing photomultiplier means for converting fluorescent energy into electrical signals; F. said movable mirror means also simultaneously directing fluorescent energy from said sample of said reference, respectively, toward said photomultiplier means; G. selectively directing said beam toward said first position or said second position, respectively, while simultaneously directing fluorescent energy from said sample or reference, respectively, toward said photomultiplier means; H. converting said electrical signals to digital signals; I. integrating said digital signals over a predetermined time interval; J. comparing the signals from said sample with the signals from said reference, thereby producing the ratio of sample to reference intensity; K. dispensing reagent into said sample to thereby change the fluorescence level of said sample relative to said initial fluorescence level; L. repeating steps (G) through (J); and M. printing out the desired information.

Claim 6

A method for high precision biochemical analysis comprising: A. providing a source of a beam of radiant energy; B. providing a movable mirror means for selectively directing said beam toward a first position and a second position, respectively when irradiated by said radiant energy; C. providing a sample of a type exhibiting an initial level of fluorescence to be measured at said first position; D. providing a reference in said second position; E. providing photomultiplier means for converting fluorescent energy into electrical signals; F. said movable mirror means also simultaneously directing fluorescent energy from said sample of said reference, respectively, toward said photomultiplier means; G. selectively directing said beam toward said first position or said second position, respectively, while simultaneously directing fluorescent energy from said sample or reference, respectively, toward said photomultiplier means; H. converting said electrical signals to digital signals; I. integrating said digital signals over a predetermined time interval; J. comparing the signals from said sample with the signals from said reference, thereby producing the ratio of sample to reference intensity; K. dispensing reagent into said sample to thereby change the fluorescence level of said sample relative to said initial fluorescence level; L. repeating steps (G) through (J); and M. printing out the desired information.