This talk will describe current and planned neutrino beams, especially the Tokai-to-Kamioka (T2K) experiment, now under construction in Japan. T2K will examine a beam of muon neutrinos produced at J-PARC on the east coast of Japan. With two neutrino detectors, one located near the origin of the beam, and another detector located 295 km away, T2K will look for the disappearance of muon neutrinos and the appearance of electron neutrinos over a long distance. T2K will begin taking data in 2009.
The protein architecture of a microtubule attachment site at the kinetochore can be revealed by answering two questions: (1) how many copies of each protein complexes exist in one kinetochore-microtubule attachment, and (2) what is the arrangement of protein complexes within the microtubule attachment site. The relatively simple kinetochore in budding yeast (Saccharomyces cerevisiae), which attaches to only one microtubule plus-end, provides an excellent model to address these questions. I have counted the copy number of each protein complex at the budding yeast kinetochore using an in vivo technique of quantitative fluorescence microscopy of GFP-tagged kinetochore proteins. To determine the relative localization of each complex within the kinetochore, I have adapted Single molecule High Resolution Co-localization (SHReC), an in vitro technique, for use in vivo. Using this technique, I am currently assembling a protein localization map for the yeast kinetochore with ~ 10 nm accuracy. Using the protein counts and localization data along with available biochemical and structural information for each protein complex, I will construct a 3-D visualization of the kinetochore-microtubule attachment. Finally, analysis of protein counts in two other fungal systems (fission yeast and Candida albicans) established that complex kinetochores with multiple microtubule attachments are built by repeating an evolutionarily conserved subunit structure that is equivalent to the budding yeast kinetochore.
Most of the core kinetochore proteins are conserved in all eukaryotes, from budding yeast to humans. Therefore study of the budding yeast kinetochore can provide the foundation for quantitative models describing the complex behavior and regulation of the human kinetochore. The 3-D protein architecture assembled in this study will provide the molecular context for future in vivo studies investigating the biophysical mechanisms underlying the basic functions of a kinetochore-microtubule attachment such as force generation, and potential factors (e.g. an opposing force) that regulate its activity.
In the second part of my talk, I will address an important phenomenon in Optical Coherence Imaging (OCI)—speckle. I will discuss the limits to the information content of OCI images of biological samples arising from speckle and scatter. The understanding of such limits is of fundamental interest, especially because the technological advances in OCI-based biomedical imaging are occurring at a rapid pace and we need to know where the limits are.
In the last part of my talk, I will introduce an effective, high-sensitive measuring platform that uses speckle as a source of information (rather than noise) for the assessment of the mechanical and dynamical properties of tissue and cells. This novel optical approach is relatively simple and yet powerful, and has the capacity to impact on global public health problems, such as malaria in resource-limited settings.
1. M. V. Goldman, D. L. Newman and A. Mangeney, PRL 99, 145002 (2007).
Contact:
Michael Hermele