Abstract: A quantitative fluorometer for multiple fluorophores having dual time-modulated beams of excitation light. Each beam is synchronized with a separate detector and lock-in amplifier. The fluorophores are simultaneously excited and the combined fluorescent emission is resolved into components corresponding to each fluorophore. Confocal scanning means are used to excite and detect fluorescent emission from locations throughout a volume. The location specific output of each amplifier is stored in a computer which resolves the emission into the components corresponding to each fluorophore. The location specific data may be further processed or visually displayed. Multiple amplifiers for each detector channel allow phase discrimination in each channel so that prompt and delayed fluorescence may be measured, allowing use of multiple fluorophores in each detector channel.

Abstract: A quantitative fluorometer for multiple fluorophores having a separate time-modulated beam of excitation light for each fluorescent target. Each beam is synchronized with a separate lock-in amplifier. The fluorophores are simultaneously excited and the combined fluorescent emission is resolved into components corresponding to each fluorophore. Confocal scanning means are used to excite and detect fluorescent emission from locations throughout a volume. The location specific output of each amplifier is stored in a computer which resolves the emission into the components corresponding to each fluorophore. The location specific data may be further processed or visually displayed.

Abstract: A method for microphotometering individual volume elements of a microscope specimen 10, comprising generating a luminous dot or cursor and progressively illuminating a plurality of part elements in the focal plane 11 of the microscope through the specimen. The mutual position between the specimen and the focal plane is then changed and a plurality of part elements in the focal plane are illuminated. Reflected and/or fluorescent light and transmitted light respectively created by the illumination is collected, detected and stored for generating a three-dimensional image of that part of the specimen composed of the volume elements. Illumination of multiples of part elements is implemented by deflecting the cursor and/or by moving the specimen. The change in the relative mutual position between the specimen and the focal plane of the microscope is effected either by displacing the specimen or the objective.

Abstract: A method of microphotometering individual volume elements of a microscope specimen 10, comprising generating a luminous dot or cursor and progressively illuminating a plurality of part elements in the focal plane 11 of the microscope through the specimen. The mutual position between the specimen and the focal plane is then changed and a plurality of part elements in the focal plane are illuminated. Reflected and/or fluorescent light and transmitted light respectively created by the illumination is collected, detected and stored for generating a three-dimensional image of that part of the specimen composed of the volume elements. Illumination of multiples of part elements is implemented by deflecting the cursor and/or by moving the specimen. The change in the relative mutual position between the specimen and the focal plane of the microscope is effected either by displacing the specimen or the objective.