Superfund Research Program
Image Analysis and Bioassays Core
Project Leader: Robert C. Burghardt
Grant Number: P42ES004917
Funding Period: 2000-2008
The Image Analysis Core continues to work closely with SBRP investigators to provide analytical microscopy and image analysis support services. Improvements in analytical microscopy instrumentation and services including specialized training programs remains an ongoing priority. During the past year new analytical instrumentation has been purchased based upon a needs-assessment survey of SBRP investigators which identified requirmements for more advanced and higher throughput laser capture microdissection technologies and whole animal imaging capabilities. Institutional support of $398,723 was obtained to purchase two instruments:
- a Molecular Devices Veritas Microdissection system and
- a Berthold NightOWL II LB 983 bioluminescence imaging and biofluorescence imaging system.
The laser capture system is equipped with epifluorescence illumination along with ultraviolet (UV) laser cutting and infrared laser (IR) capture capabilities that has improved speed, precision while maximizing biomolecule integrity needed for SBRP applications. The NightOWL system is being used to provide real-time imaging to non-invasively monitor and record cellular and genetic activity within living organisms. During the current funding period, the Endocrine Disruptors project and the Genotoxicity of Complex Mixtures project have continued to make use the entire range of vital imaging instruments (whole animal, live cell and digital imaging workstations, confocal and multiphoton microscopes) and applications for routine bioassays of cellular function in toxicant-exposed cell types including: the FRAP approach to quantify effects of PAH mixtures on intercellular communication (a tumor promoter assay) and assessment of cytochrome P450 enzyme induction (EROD assay); single-cell and bulk analysis of cytochrome P450 enzyme induction; reactive oxygen species production, apoptosis assays, and analysis of intracellular Ca2+ homeostasis and Ca2+ waves and oscillations. The Endocrine Disruptors project also continued utilization of immunocytochemistry as well as direct analysis of estrogen receptor (ER), aryl hydrocarbon (AhR), Sp1-Sp4 and coactivator protein-protein interactions using FRET assays. Other analyses involved the evaluation of the efficacy of specific siRNA constructs on the expression and function of these proteins. The Genotoxicity of Complex Mistures project continues use of electron microscopy to monitor ultrastructural changes and apoptosis in neural tubes of fetal mice exposed to arsenic.