We request funding for a newly developed instrument called a bio-imaging analyzer (also known as a phosphorimager) that can be used for the detection of radioactively labeled molecules. Isotopes such as 32p, 125I, 35S, and 14C, that are widely used in the study of proteins, nucleic acids, and other macromolecules can be detected using this instrument. Although autoradiography with X-ray film has been used traditionally for such applications, the bio-imaging analyzer is superior in that: (1) it is up to 100 times more sensitive than X-ray film; (2) it on average requires 10-fold lower exposure times; (3) it possesses a much broader dynamic range for comparing two samples of greatly different radioactive intensity than X-ray film; (4) it provides the opportunity for rapid and direct quantitation of data by computer-based methods; and (5) it can be used in combination with newly developed software to read samples such as DNA sequencing gels automatically. The bio-imaging analyzer will be used by twenty four labs at Caltech consisting of approximately three hundred researchers, including postdoctoral fellows, graduate students, and technicians. The instrument will be used in combination with a wide variety of techniques, including: immunoblotting; Northern and Southern blotting; CAT assays; RFLP analysis; DNA sequencing; two-dimensional gel analysis; phosphopeptide mapping; and PCR techniques. In these various applications, the analyzer will be used: (1) for quantitation of data; (2) for image processing and computer analysis of data; and (3) for automated collection of data. Given its versatile capabilities, the bio-imaging analyzer should greatly enhance our research efforts, both quantitatively and qualitatively.
