Method and apparatus for the measurement of low-level laser-induced fluorescence

An apparatus for measuring the excited fluorescence of an object having a binding fluorescent dye application comprising:

• means for generating a coherent beam of light;
• means for expanding said coherent beam of light;
• means for focusing said expanded beam of light;
• a three dimensional scanner having means for securing such object in a fixed position, said scanner receiving and passing said focused beam of light over such fixed object in a selected pattern and at a selected rate, said scanner having X, Y coordinate scanning capability with simultaneous Z coordinate correction to cause said focused beam of light to have a topographical field of focus in the plane of said object;
• means for controlling the pattern and rate at which said beam of light is passed through said translucent object;
• a biased cut optical fiber faceplate for receiving and directing the fluorescent light transmitted from said object;
• means for filtering said fluorescent light emanating from said fiber faceplate;
• means for diffusing said filtered light;
• means for collecting and measuring the intensity of the fluorescent light transmitted through said object;
• and
• means for sequentially recording data regarding the intensity of the fluorescent light transmitted from said object as a function of the location of said scanning beam of light on said object.

An apparatus for measuring the excited fluorescence of an object having a finding fluorescent dye application comprising;

• a laser for generating a coherent beam of light at a specified wavelength with the range of 350 to 540 nanometers;
• a variable beam expander for expanding said coherent beam of light;
• a focusing lens to focus said expanded beam of light;
• a three dimensional scanner having means for securing such object in a fixed position, said scanner receiving and passing said focused beam of light over such fixed object in a selected pattern and at a selected rate, said scanner having X, Y coordinate scanning capability with simultaneous Z coordinate correction to cause said focused beam of light to have a topographical field of focus in the plane of said object;
• means for controlling the pattern and rate at which said focused beam of light is passed over said object;
• a biased cut optical fiber faceplate for receiving and directing the fluorescent light transmitted from said object;
• filter means for receiving said fluorescent light from said fiber optic bundle and removing unwanted light input;
• diffusion means for diffusing said filtered light;
• a photomultiplier for measuring the intensity of the fluorescent light transmitted and received from said diffusion means;
• and,
• means for sequentially recording data regarding the intensity of the fluorescent light transmitted from said object as a function of the location of said scanning beam of light on said object.