AN IMAGE ACQUISITION AND PROCESSING SYSTEM FOR THE ANALYSIS OF FLUORESCENCE FROM STAINED DNA GELS

EVALUATION OF DAMAGED DNA COMPONENTS IS NORMALLY PERFORMED VIA AGAROSE GEL ELECTROPHORESIS OF DNA SAMPLES STAINED WITH FLUORESCING DYE. ALTHOUGH THIS METHOD PROVIDES ADEQUATE SENSITIVITY AND RESOLUTION IN THE IDENTIFICATION OF DNA COMPONENTS, THE LACK OF AVAILABLE INSTRUMENTATION TO CONVERT THE FLUORESCING IMAGE INTO A DIGITIZED RECORD FOR COMPUTER ENTRY AND THE LACK OF ANY TECHNIQUE FOR STANDARDIZING ANALYSES PERFORMED UNDER VARYING CONDITIONS SEVERELY LIMIT THE USEFULNESS OF THIS ANALYSIS. THE GOAL OF THIS PROJECT IS TO DEVELOP AN INSTRUMENT THAT MAKES IT POSSIBLE TO DIGITIZE STAINED DNA GELS DIRECTLY INTO ANY COMPUTER (VIA A STANDARD COMMUNICATIONS INTERFACE), STANDARDIZE THE RESULTS, AND PERFORM STATISTICAL ANALYSIS OF THE DATA. THIS INSTRUMENT WILL QUICKLY SCAN THE GEL USING AN OPTICAL SYSTEM THAT HAS HIGH PHOTOMETRIC RESOLUTION AND VERY HIGH SPATIAL PRECISION. THE DIGITIZED DATA WILL THEN BE ELECTRONICALLY PREPROCESSED (BACKGROUND SUBTRACTION, FILTERING, DATA COMPRESSION, ETC.) TO PROVIDE CORRECT, NOISE-FREE DATA STRUCTURED FOR STORAGE WITH MINIMAL MEMORY REQUIREMENTS. THE STORED DATA WILL THEN BE AVAILABLE FOR USE IN IMAGING THE GEL, PRODUCING STATISTICAL COMPARISONS, AND GENERATING GRAPHIC DISPLAYS. THE DATA WILL BE ARCHIVABLE USING MEDIA SUCH AS MAGNETIC TAPES OR DISKS. THIS INSTRUMENT WILL OFFER A SOLUTION TO PROBLEMS LONG ASSOCIATED WITH AGAROSE GEL ANALYSIS OF DNA. THE DIRECT QUANTIFICATION OF FLUORESCENCE IN STAINED DNA GELS WILL ALLOW IMMEDIATE ACCESS TO THE DATA BY A COMPUTER, INSTANTLY PROVIDING SUCH INFORMATION AS AVERAGE MOLECULAR WEIGHT. THE INSTRUMENT WILL ENABLE DIRECT COMPARISON OF SEPARATE ANALYSES PERFORMED UNDER VARYING CONDITIONS, ALLOWING FOR COMPLETE NORMALIZATION OF RESULTS AND GENERATION OF STATISTICAL DATA INVOLVING MANY INDIVIDUAL TESTS.