CLINICAL APPLICATION OF A LASER SCANNING CYTOMETER

WE HAVE DEVELOPED A COMPUTER CONTROLLED LASER SCANNING CYTOMETER POTENTIALLY CAPABLE OF MEASURING MULTIPLE WAVELENGTH FLUORESCENCE AND MULTIPLE ANGLE SCATTER OF CELLS ON A SOLID SUPPORT SUCH AS A SLIDE, CUVETTE, OR CULTURE PLATE AT RAPID ENOUGH RATES TO PROCESS 10,000 CELL SAMPLES IN TWO MINUTES. IN ADDITION TO THE STANDARD FLOW CYTOMETRY MEASUREMENTS, THIS INSTRUMENT INCLUDES IN ITS DATA LISTS AND IT DISPLAYS INTERCHANGEABLY 1) COURSE MORPHOLOGICAL PROPERTIES, 2) THE EXACT TIME OF MEASUREMENT AND 3) CELL POSITION. SINCE CELL POSITION IS RECORDED FOR EACH CELL, CELLS CAN BE VISUALLY OBSERVED DURING RUNS OR RETROSPECTIVELY, AND CELLS CAN BE REMEASURED MULTIPLE TIMES TO FOLLOW KINETIC PROPERTIES OF SELECTED SUBPOPULATIONS. WE HAVE BUILT TWO DUAL PARAMETER INSTRUMENTS, AND WRITTEN SOFTWARE TO CONTROL THEM AND DISPLAY RESULTS. WE WILL DEVELOP AND EVALUATE TECHNIQUES FOR MEASURING CLINICAL SPECIMENS OF HUMAN CANCER WITH THEM AS A POSSIBLE AID IN DIAGNOSIS AND IMPROVED PROGNOSIS AND TREATMENT. THIS REQUIRES THAT WE SENSITIVELY MEASURE FLUORESCINATED NUCLEIC ACID PROBES OR ANTIBODIES AND SIMULTANEOUSLY AND PRECISELY MEASURE THE FLUORESCENCE OF A DNA STAIN SUCH AS PROPIDIUM IODIDE. PHASE I AIMS ARE TO TEST THE FEASIBILITY OF 1) DUAL FLUORESCENCE MEASUREMENTS ON CLINICAL SPECIMENS OF HUMAN TUMORS IN COMPARISON WITH CONVENTIONAL FLOW CYTOMETRY, AND 2) IMAGING CELLS OR FIELDS AS THEY ARE SCANNED, A) TO CONVENIENTLY SELECT THOSE TO BE MEASURED, OR B) TO VIEW CELLS SELECTED BY THE INSTRUMENT FOR VISUAL CLASSIFICATION.