The objective of this laboratory is to develop techniques for quantitative 3D light and electron microscopy and to make these available to the scientific community for research and education. A 400 kV transmission electron microscope allows examination of specimens 10-20 times thicker than usual and has been modified for an unlimited range of specimen tilt angles for 3D imaging. A laser scanning confocal light microscope provides optical sectioning in thick specimens, including wet and living specimens, superior to that obtainable using ordinary light microscopes. The foci in the core research are quantitation, comparison of images using different imaging modalities, and the evaluation of the effects of preparation methods and other potential sources of artifacts. Images are processed using image analysis programs imported from other sources or developed here. Programs are available for reconstruction from serial section images and from images taken at various tilts (including stereo images). These programs provide 3D localization of particulate markers (gold particles), reconstruction of membranes and other surfaces, and quantitative analysis of networks. Initially, manual processes are implemented. We plan in the longer term to automate as many processes as possible. Current collaborative research projects are focussed on the localization of surfaces and surface receptor molecules in motile cells, the organization of the cytoplasmic matrix, the structure of protein networks, the structure of the transverse tubular system in skeletal muscle, and the development of the fibrillar system in differentiating cardiac and skeletal muscle cells. Service projects cover a broad range of topics. Users have come from several surrounding states and from Japan. Training is done on-site at the facility, and through workshops presented at the facility and elsewhere. Dissemination of information on the facility is done through publications, lectures and, posters presented at national and international meetings, and direct mailing to biomedical scientists.

Agency
National Institute of Health (NIH)
Institute
National Center for Research Resources (NCRR)
Type
Biotechnology Resource Grants (P41)
Project #
5P41RR002483-09
Application #
2281648
Study Section
Special Emphasis Panel (SSS (P4))
Project Start
1984-12-01
Project End
1997-08-31
Budget Start
1995-09-15
Budget End
1996-08-31
Support Year
9
Fiscal Year
1995
Total Cost
Indirect Cost
Name
University of Pennsylvania
Department
Biology
Type
Schools of Arts and Sciences
DUNS #
042250712
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Deng, Manqi; Williams, Carmen J; Schultz, Richard M (2005) Role of MAP kinase and myosin light chain kinase in chromosome-induced development of mouse egg polarity. Dev Biol 278:358-66
Tutuncu, Levent; Stein, Paula; Ord, Teri S et al. (2004) Calreticulin on the mouse egg surface mediates transmembrane signaling linked to cell cycle resumption. Dev Biol 270:246-60
Brown, Rebecca L; August, Shelley L; Williams, Carmen J et al. (2003) AKAP7gamma is a nuclear RI-binding AKAP. Biochem Biophys Res Commun 306:394-401
Deng, Manqi; Kishikawa, Hidefumi; Yanagimachi, Ryuzo et al. (2003) Chromatin-mediated cortical granule redistribution is responsible for the formation of the cortical granule-free domain in mouse eggs. Dev Biol 257:166-76
Brown, Rebecca L; Ord, Teri; Moss, Stuart B et al. (2002) A-kinase anchor proteins as potential regulators of protein kinase A function in oocytes. Biol Reprod 67:981-7
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Travis, A J; Merdiushev, T; Vargas, L A et al. (2001) Expression and localization of caveolin-1, and the presence of membrane rafts, in mouse and Guinea pig spermatozoa. Dev Biol 240:599-610
Stein, P; Schultz, R M (2000) Initiation of a chromatin-based transcriptionally repressive state in the preimplantation mouse embryo: lack of a primary role for expression of somatic histone H1. Mol Reprod Dev 55:241-8
Li, J; Ashton, F T; Gamble, H R et al. (2000) Sensory neuroanatomy of a passively ingested nematode parasite, Haemonchus contortus: amphidial neurons of the first stage larva. J Comp Neurol 417:299-314

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