2 years ago

2004 Scientific Report

Transcription factor

Transcription factor expression in human cells This is a confocal, fluorescent image of a unique transcription factor’s expression in human cells grown on cover slips. The blue stain labels the DNA of the cell and defines the cell nucleus; only the nucleus is seen in this image. The nucleus is the site of transcription, an activity in which the DNA genetic message is read and processed. A well-characterized transcription factor complex is labeled with red stain. The green stain marks the novel protein believed to be a component of the transcription complex. The green pixels are all within the nucleus and co-localize with many of the red pixels. The green pixels occupy a unique globular, peri-nuclear space. This is morphological correlation of a functional association between the novel protein and the transcription complex. Analysis indicates that 19.41% of the red pixels are co-localized with green pixels. This quantification is consistent with the novel green protein forming a portion of the transcription complex. (Resau; quantification by Kort. This work is part of a collaboration between Dr. Resau and Dr. O. Rosen of Harvard University.) 38

Laboratory of Analytical, Cellular, and Molecular Microscopy and Laboratory of Molecular Diagnostics James H. Resau, Ph.D. Dr. Resau received his Ph.D. from the University of Maryland School of Medicine in 1985. He has been involved in clinical and basic science imaging and pathologyrelated research since 1972. Between 1968 and 1994, he was in the U.S. Army (active duty and reserve assignments) and served in Vietnam. From 1985 until 1992, Dr. Resau was a tenured faculty member at the University of Maryland School of Medicine, Department of Pathology. He retired from the University of Maryland and joined the National Cancer Institute. Dr. Resau was the Director of the Analytical, Cellular and Molecular Microscopy Laboratory in the Advanced BioScience Laboratories–Basic Research Program at the National Cancer Institute–Frederick Cancer Research and Development Center, Maryland (1992–1999). He joined VARI as a Special Program Senior Investigator in June 1999, and in 2003 he was promoted to Deputy Director. In 2004, Dr. Resau assumed the direction of the Laboratory of Microarray Technology to consolidate the imaging and quantification of clinical samples in a CLIA-type research laboratory program. Staff Robert Sigler, D.V.M., Ph.D. Eric Kort, M.S. Bree Berghuis, B.S., HTL (ASCP), QIHC Pete Haak, B.S. Eric Hudson, B.S. Paul Norton, B.S. J.C. Goolsby Laboratory Members Brandon Leeser Christine Moore Amy Percival Students Rebecca Roe Huang Tran Research Interests Our laboratories are responsible for producing and interpreting images and cDNA microarrays as they relate to the diagnosis and characterization of disease, injury, and differentiation. Although primarily we study cancer, we work with a variety of diseases. The ACMM lab works closely with VARI investigators, as well as with collaborators from outside the Institute, to provide support for a variety of imaging and pathology projects. We have a special interest in the quantification of imagery. We have two confocal microscopes that enable us to visualize organelles and proteins in cells or tissues. We have studied the location of two gene-targeted proteins (GFP and RFP) within a cell in three dimensions, using them with a nuclear (DAPI) marker. We have integrated laser-capture microdissection instrumentation into our imaging program, as well as paraffin- and frozen-section staining. We also provide histotechnology services, histopathology consultation, immunostaining expertise, and direction of the human tissue services. The MD lab produces highquality cDNA microarrays in accordance with CLIA protocols for use in the study of tissues, lesions, and organisms. Our laboratory is primarily responsible for the archived clinical histopathology program called SPIN (Frostbite). This program allows investigators to use existing clinical samples to assess the expression of proteins in human disease samples. In addition, we use these blocks to prepare a wide variety of tissue microarrays for research purposes. Currently the archive holds approximately 150,000 tissue samples/paraffin blocks. They are not directly linked to any personal identifiers or names and meet HIPAA/CLIA regulations. The material from future years will be available with digital information on age, sex, and diagnosis and linked to image files. These samples will be used in cellular and molecular protocols approved by our Institutional Review Board (IRB). The samples and demographics are identified with basic information in a webbased, interactive format for determination of diagnosis, prognosis, and therapy. In the first years of operation for SPIN there have been 24 users registered who have submitted 508 requests for searches and 75 subsequent tissue requests. In collaboration with George Vande Woude and Rick Hay of VARI, we have augmented this program with the collection of freshly frozen human 39

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