Cell Isolation and Systems Analysis

Additional Information

The Cell Isolation & Systems Analysis (CISA) capability at EMSL includes techniques for isolating cells from complex cell populations or environmental samples for further 'omics and imaging analyses. EMSL specializes in quantitative live cell fluorescence imaging with single molecule sensitivity, super resolution fluorescence and electron microscopy techniques, and transcriptomics and proteomics analyses. These capabilities provide the foundation for attaining a molecular-level understanding of protein functions and microbial-community dynamics to enable molecular level investigations of single cells and the pursuit of systems biology.

Capability Detail

Bioreactors and Cell Culture – EMSL provides instrumentation and expertise for the controlled growth of diverse microbial cell types, at scales ranging from 100 µl wells to multi-liter reactors, which can be used in downstream analysis in a wide array of EMSL instruments. The Bioscreen-C instrument allows the automated, real-time growth rate analysis of up to 200 independent wells for determining the effect of media composition on the growth of microbial cells. The Micro-24 bioreactor consists of 24 independently controlled reactors (5-7 ml cultures) to support the growth of microorganisms. Each reactor cassette is independently monitored and controlled for dissolved oxygen levels, pH, and temperature, with real-time monitoring of all parameters. The BioFlo 310 benchtop fermentor-bioreactor system can regulate up to 32 parameters in each fermentor, enabling the integration and control of multiple analyzers, pumps and sensors. In addition, a light-cycling growth chamber is available to support the growth of cyanobacteria, and a mammalian cell culture facility is available to support the growth of human and other mammalian cells.
Cell and Organelle Isolation – Resources are available at EMSL for detecting and isolating distinct cells and organelles for further single-cell and other analyses. The Influx, a one-of-a kind flow cytometer/cell sorter, provides powerful detection capabilities and diverse sorting approaches, incorporating cutting-edge technologies to support new sorting modes optimal for complex cell populations. For example, users can sort cells by their proportional DNA content, hybridized mRNA content, and specific protein or molecule content. Cells can be sorted in a two-dimensional pattern that replicates the pattern on a bivariate dot plot, enabling subset identification in complex mixtures of cells. Multiple collection options, including sorting directly into micro-titer plates, slides, or Petri dishes, enable sorting for single-cell cloning and single-cell transcriptomics or imaging. Importantly, Influx technology supports the detection and sorting of nano-scale particles, making it highly suitable for the sorting and analysis of organelles and microorganisms. EMSL also provides a Laser-Capture Micro-Dissection microscope, equipped with 100 x objective lens for enriching distinct organelles, or isolating single cells from cell populations or tissue sections for further analyses.
Fluorescence Microscopy/Spectroscopy – A suite of non-conventional, cutting-edge, high-spatial and temporal resolution microscopes, including super resolution and single-molecule fluorescence microscopy, are available at EMSL, allowing users to gain new insights into living cells. EMSL's Multi-Photon microscope seamlessly integrates nonlinear two-photon excitation, laser-scanning confocal microscopy, and fluorescence lifetime imaging (FLIM) in one fully automated upright system. Nonlinear two-photon excitation allows minimally invasive and deep-penetrating, high-resolution 3D imaging of living systems, and FLIM supports quantitative fluorescence resonance energy transfer (FRET) for investigating molecular interaction dynamics in living cells. In addition to the scanning confocal microscope, the system also supports DIC imaging. EMSL has developed and offers STORM (stochastic optical reconstruction microscope), also known as PALM (photo-activated localization microscopy) – a fully automated super-resolution fluorescence microscope that enables fluorescence imaging of the intact cell with nanometer resolution. STORM or PALM, incorporates single-molecule fluorescence techniques to reconstruct super-resolution images using software that automatically acquires and analyzes data in real time. Structured illumination fluorescence microscope (SIM) is now also available for super resolution fluorescence imaging with 100 - 130 nm resolution in 3D. The fluorescence SIM enables the imaging of any fluorescent dye or protein. High-sensitivity total internal reflection fluorescence (FLIM) microscopes are also available at EMSL to support time-lapse single-molecule fluorescence imaging of individual proteins or organelles in living cells.
Electron and Ion Microscopy – EMSL's Microscopy Capability provides leading-edge, high-resolution transmission and scanning electron microscopes, such as the CRYO 2005 and the Dual-beam FIB/SEM microscopes, and helium ion microscope for unprecedented resolution and focal depth.
Transcriptomics – EMSL offers two Applied Biosystems SOLiD platforms for massively parallel next-generation sequencing. The platforms can be used for a wide array of applications, such as global gene expression analysis with single base resolution, single-cell or meta-transcriptomic analysis, regulation of gene expression including ChIP-Seq or microRNA analysis, and the identification of novel genes or isoforms. The platforms offer a non-biased, highly quantitative, and accurate method to sequence DNA or RNA from any source and supersede that of previous technologies. Multiple unique samples can be sequenced in parallel in a single run with high coverage using barcoding, saving time and reagents. Because of a novel two-base encoding method that discriminates between actual single nucleotide polymorphisms (SNPs) and sequencing errors, sequencing from normal or mutated cells can identify gene expression or epigenetic (gene methylation analysis) alterations as well as SNPs in a quantitative and accurate manner. In addition to transcriptomics analysis, proteomics analysis is provided by Mass Spectrometry Capability, and metabolomics analysis by the NMR Capability. Together, EMSL's powerful resources and expert staff allow researchers to conduct parallel analyses, using a systems biology approach, under one roof.

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