The expertise and instrumentation to perform most flow cytometric assays that have been described in the literature is available within the expertise of the collective personnel and the physical resources of the Robert H. Lurie Comprehensive Cancer Center Flow Cytometry Core Facility. One of the missions of the Core Facility is to provide access to the breadth of more complex cytometry assays that investigators would not otherwise have and to work collaboratively to bring any assay needed on line. The services provided fall into several categories, the total of which illustrates the value and extent of services offered to the community of NU researchers.
The assays offered by the facility range from routine cell cycle analyses and immunophenotyping to complex multi-laser applications and high speed cell sorting. Examples of the assays available include, but are not limited to the following:
- DNA content/cell cycle measurement, including the simultaneous measurement of multiple fluorescent probes if required.
- Immunofluorescence analyses (three to six color analyses are routinely performed, greater than seven color are under development) including characterization of plasma membrane antigens/receptors, cytoplasmic proteins, and nuclear antigens.
- Characterization of cell populations based on scattered light intensity measurements, and autofluorescence.
- Cell sorting including viable, sterile cell sorting and spot blot analyses.
- Intracellular calcium flux
- A range of apoptosis assays including Annexin V, TUNEL, mitochondrial membrane potential, and measurement of proteins involved in apoptosis regulation such as multiple members of the BCL-2 family of proteins, CD95 and caspase protein levels/activity
- Fluorescence Resonance Energy Tranfer (FRET)
- Nanoparticle characterization
- Bivariate and univariate chromosome analysis
- Receptor-ligand interactions
- Cell proliferation studies including BrdU incorporation and CFSE tracking
- Viability assays (membrane exclusion and metabolic viability).
- Various function assays including oxidative metabolism, neutrophil function (oxidative burst, phagocytosis), cytoplasmic pH, membrane potential.
- Kinetic analyses
- Signal transduction pathway analyses (simultaneous assessment of multiple intracellular phosphorylated epitopes combined in complex multi-color assays)
- Sample preparation and staining
Full service: consultation and training
The Lurie Flow Cytometry facility is unique in the fact that it offers investigators the entire spectrum of cytometric experiment management, if desired, all the way from initial design consultation to the creation of graphics for publication. It is entirely up to the investigator what level of involvement the facility staff takes. The staff will prepare samples including staining, data collection, quality control, data analysis/interpretation, and creation of graphics. Alternatively, if the investigator chooses the facility will solely provide consultation on any of the above so that the research is entirely in the hands of the investigator. Sample preparation provided by the flow lab personnel has proven to be extremely valuable in that a subset of investigators may lack technologist time, or expertise, to carry out properly designed flow experiments. This service has been particularly helpful to more clinical orientated investigators allowing expansion of their laboratory based studies.
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