Today is Thursday, Apr. 19, 2018

Department of Environmental Health

Center for Environmental Genetics

Confocal Imaging and Immune Monitoring Cores (formerly the Flow Cytometry Core)

The Confocal Imaging and Immune Monitoring Cores were introduced in 2015. Two key facility upgrades are the Zeiss Confocal microscope LSM 700 and ImmunoSpot® Analyzer.

Current research on environmental toxicants on single cells is moving to include RNA and DNA sequence analyses, as well as imaging of DNA structural states of the epigenome, RNA transcription, chromosomal conformation, DNA methylation, open chromatin and small-molecule metabolome assays. Using the confocal microscope for visualization of sufficient variables within each cell's tissue context regardless of molecule type (protein, RNA, DNA etc.) will give a true picture of the multidimensional spatial dynamics of the cellular ecology. Such measurements will dissect the molecular building blocks upon which biology occurs and will lead to a fuller understanding of biological processes. This confocal microscope will generate high-resolution, high quality, microscopy-based images. It may also be useful for isolation of single cells in conjunction with (prior to/to verify) RNA-sequencing, hence its location close to the Genomics, Epigenomics and Sequencing Core (GESC).

ImmunoSpot® Analyzers are designed for scanning and evaluating single cell-based spots that reflect the protein expression, function, and growth of cells. Technically, the ImmunoSpot® S6 Micro’s zoom lens permits seamless switching from 6-, to 384-well plates and can zoom in for counting objects <10 µm in size – including individual cells. It is a low maintenance and high performance instrument for measuring the functional responses of and clustering growth derived from single cells at a high-throughput plate-based setting. The ImmunoSpot® Analyzer can be applied to, but not restricted to, the following CEG-related or supported studies involving environmentally induced diseases and conditions: (i) characterization the production of cytokine, surface markers, or soluble molecules from rare cancer stem cells with high sensitivity and accuracy to capture individual cells; (ii) functional measurement of the cytokine production from rare immune cell subsets, e.g., regulatory T cells, anti-inflammatory immune cells, etc., from human samples in response to toxicant-treatment or environmental exposure; (iii) many kits have been commercially developed for testing cytokine production that can be conveniently applied in toxicology studies;  (iv) the growth of microbes from various pathogenic sources or from complex microbiota pools can be recorded, quantified, and differentiated using the high-resolution camera and analytical software, which can be applied for studying the environmental toxicity on microbiota.The ImmunoSpot® Analyzer is highly advantageous in functional characterization of rare cells from limited human samples and in the survey of rare subsets from exposure-related epidemiology studies using high-throughput microplate analyses

 Research supported by the Center for Environmental Genetics should  NIEHS P30-ES006096