Stowers
Microscopy Center
Head: Winfried Wiegraebe
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Search Stowers Microscopy Center and other Stowers Research Web Pages:Place service request (SIRIS), for internal use only.Make microscope reservation (LIMS), for internal use only. Projects
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We have set up a system to image the inside of the eye of living mice. It is based on a slit-lamp SL 120 from Carl Zeiss Meditec. Data acquisition is done by a Carl Zeiss Axiocam HR attached to the microscope using a universal video adapter manufactured by TTI Medical. To image the anterior chamber we use a goniolens optimized for mice by Ocular Instruments as described in R.S.Smith et al. James P. Gilman describes on The Ophthalmic Photographers' Society web page how to use a slit lamp for biomicrography. |
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The brain of living drosophila is a powerful model system to study neural circuits and memory formation. The large number of genetic tools available allow for modifications of biochemical properties. The small size makes imaging less daunting. On the other hand, the small size makes handling and sample preparation of the brain more challenging. The fly brain is infamous for it's strong auto-fluorescence. The cuticle makes imaging without opening the head nearly impossible. We are tackling these challenges using two photon microscopy (NLO) and linear unmixing. |
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Gene transcription by RNA polymerase II, responsible for the production of cellular mRNA, can be a critical target for regulating which complement of the cellular genome is expressed. An understanding of the mechanisms underlying the various stages of transcription will be crucial to being able to find solutions to the problems caused by misexpression of genes, problems that may give rise to a host of human diseases. Regulation of RNA polymerase II activity can occur both during the initiation and elongation stages of transcription. A protein which affects transcription elongation of RNA polymerase II, eleven-nineteen lysine-rich in leukemia (ELL), was initially identified as a gene fusion partner of the mixed lineage leukemia (MLL) gene in some patients with acute myeloid leukemia. In vitro studies have shown that the ELL protein is capable of interacting with RNA polymerase II and increasing its catalytic rate of transcription elongation. ELL has been found associated with several proteins. Simone et al. identified EAF1 (ELL-associated factor 1) via a yeast two-hybrid screen with ELL. As with the MLL-ELL gene-fusion, an MLL-EAF1 gene fusion is able both to transform primary cells, and to induce a leukemia state in mice when expressed in injected bone marrow cells. Although ELL and RNA Polymerase II have been shown to coimmunoprecipitate, it is not clear whether ELL might be recruited to RNA Polymerase II during initiation or elongation and whether the proteins remain bound during elongation. The role of EAF1 is also unclear, and although an ELL/EAF1 heterodimer affects RNA polymerase II activity, it is not clear whether ELL and EAF1 interactions are concurrent with those of ELL and polymerase. I will use fluorescence correlation spectroscopy to determine when fluorescent labeled ELL and EAF1 are recruited to the transcription complex during an in vitro transcription assay. |
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For comments and additional information, please contact Winfried Wiegraebe. Red links are for internal use only. Sorry!
