The Flow Cytometry Core lab, located in W290 at The Research Institute at Nationwide Children's Hospital, offers several platforms on which to do flow analysis and/or cell sorting. Depending on the complexity of the experiment involved there are several "levels" of cytometer available to address your specific needs. In addition, Flow Jo analysis software is available on standalone computers on both Mac and PC platforms. Flow cytometry is continually expanding its applications as a research tool. Contact us to discuss what flow cytometry can do to help facilitate your research.
Flow Cytometry has a diverse array of applications and The Research Institute at Nationwide Children's seeks to help researchers with any application they wish to pursue. As science evolves, so does the scope of flow cytometry and the facility strives to stay abreast of the latest techniques. The knowledgeable staff are here to assist you with experiment design, data acquisition, data analysis and any other questions you may have.
For the convenience of investigators at Nationwide Children's, the Flow Cytometry Core is now offering operator-assisted services and consultation. These services are perfectly geared for pilot experiments initiated by Investigators who are unfamiliar with flow-based technologies, or Investigators doing the rare flow experiment who simply wish to save time. These services are not meant to replace the regular training, education, troubleshooting and assistance that we happily provide all users, but rather, they are optional, charge-based services of convenience.
Flow based technologies are tremendously useful for the analysis of cell cycle activity (by DNA content), proliferation (Ki67), stages of apoptosis or necrosis (Annexin V combined with 7AAD) and intracellular signaling (Ca++ flux). In addition, cells carrying a certain functional significance (primitive stem cells) can be identified using reporter dyes that measure the activity of specific proteins (for instance Alkaline Phosphatase or ABC transporters).
Both the analyzers and sorters in our facility have configurations that are optimized for multicolor flow applications, and have been used successfully for the identification and phenotyping of rare cell types. These include, but are not limited to, virus-specific cytotoxic T cells, antigen-loaded dendritic cells, primitive stem cells and hematopoietic progenitors. We are continually testing the limits of these machines for their sensitivity, accuracy and consistency. Our working configurations are modern, and evolve alongside newly available technologies and dyes.
Flow cytometry is ideal for the quantification of reporter gene activity, transfection or transduction efficiency using fluorescent reporter dyes (GFP, RFP, YFP, etc) as a readout. Such an approach may be useful for the construction and optimization of fluorescent viruses, gene transfer approaches, or growth conditions for lineage-specific cell types. Our expert staff is familiar with such approaches, and can assist you in the assessment of cell viability and autofluorescence, which are often vital for accurate interpretation of results.
Numerous flow-based methods are now available for the accurate detection and quantification of cytokines and other important analytes. Intracellular cytokine staining can be used to directly visualize and phenotype cytokine producing cells on a per cell basis (like dendritic cells, macrophages or T cells), while bead-based arrays can be used to quantify multiple analytes simultaenously. For the latter, the Bio-Plex system and CBA-kits (BD Biosciences) are two such options that can be performed in our core. The Flow Cytometry Core also houses a Wallac 1620 plate reader, useful for the traditional ELISA method of cytokine detection.
Cell biologists rely heavily on flow-based approaches to study various cell activities. These include: cytotoxic T cell killing, degranulation, cellular proliferation, phagocytosis, antigen processing, migration and even immunomodulation. If functional assays are of interest, please ask us how we might assist you. We may even offer suggestions for developing novel tests suited to your research.
Cell sorting can be used for a wide range of downstream applications, such as the isolation of reporter dye+ cells (single color) or the isolation of multiple cell populations from a single sample (peripheral blood T cells, B cells, and monocytes, for example). The Flow Cytometry Core does routine testing of our sorters for performance, by assessing purity, yield, and viability of sorted cells. We also test the limitations of our machines when challenged with limiting sample sizes, the sorting of rare populations, or sorting for downstream applications involving DNA or RNA isolation steps.
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