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|  University of Hawai'i Manoa |
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| University of Hawai'i Manoa |
The of the goal of the Immunology Core of MCI, which functions as a fee-for-service facility, is to provide the Hawaii community with a service-oriented facility to meet the needs in the FACS research areas. We are equipped with BD FACSAria, BD FACSCalibur, GuavaEAsyCytePlus, Luminex 200, Beckman-Coulter ViCell, CTL-ImmunoSpot® S5 Core Analyzer, Bio-Rad and BioTek ELx808 ELISA readers and washers, Faxitron compact 43855D x-ray irradiation system, and Faxitron x-ray CP160 for in vivo animal work.
Member:
Gurary, Alexandra
Role:
Specialist, Flow Cytometry Facility
"The BD FACSAria flow cytometer is the first fixed-alignment benchtop high- speed cell sorter. The instrument can be operated at varied pressures and can acquire up to 70,000 events per second. With its fixed-optics design and digital electronics, the BD FACSAria flow cytometer enables multicolor analysis of 7 fluorescent markers and two scatter parameters at a time."
"The BD FACSCalibur™ platform offers a unique modular approach to flow cytometry and allows users to perform both cell analysis and cell sorting in an innovative single benchtop system."
"The Vi-CELL Series Cell Viability Analyzers provide an automatic and cost effective means to perform the trypan blue dye exclusion method."
"The S5 Core is CTL’s dedicated model that has been fine tailored for ELISPOT work in the 96 well format. Its features high end optics and cameras – permitting to counting objects as small as >25 µm in size which can be critical for certain ELISPOT assays, including TNF and perforin measurements. The S5 Core ideally lends itself for GLP and high-throughput upgrades. The S5 Core instrument is ideally suited for users who do one particular type of assay while having the highest expectations for image and analysis quality."
"Automated multicolor detection and absolute counts for fast and easy identification of cell phenotype and function."
"The Luminex 200 System is a flexible analyzer based on the principles of flow cytometry. The system enables you to multiplex (simultaneously measure) up to 100 analytes in a single microplate well, using very small sample volumes. The system delivers fast and cost-effective bioassay results on many assay formats including nucleic acid assays, receptor-ligand assays, immunoassays and enzymatic assays."
"FACS is used to study DNA cell content. Propidium iodide (PI) and Hoechst dyes bind to DNA and become fluorescent. PI cannot enter live cells and is included in immunofluorescent staining protocols to identify dead cells. Some Hoechst dyes can enter live cells. DNA staining can be used to study the cell division cycle. Relative DNA content shows the proportion of cells in G1, G2 and S phases. Apoptotic cells show characteristic smear on DNA staining. DNA staining examples:
* Studying changes in cell cycle in mutant cell
* Measuring apoptosis in cells after irradiation"
"FACS can be used to investigate cell biology. Calcium flux can be measured using Indo-1 markers. This can be combined with immunofluorescent stain. For example, identify T cell subpopulations by immunofluorescence and measure calcium flux in response to an activating signal. Rhodamine-123 stains mitochondrial membranes is used to measure cellular activation. Rhodamine-123 is rapidly pumped out of some cells (for example hematopoietic stem cells)."