Immuno/BioSpot Core
Cell Biology
Locations:	Factor Building 12-541 Los Angeles, CA, 90095 BSRB 188F
Email:	brgordon@ucla.edu
Phone Number:	310-206-8252
Description:	Related Resources & Cores Mucosal Immunology Core Flow Cytometry Core Laboratory Christel Uittenbogaart, Director Brent Gordon, Core Manager Services: The Immuno/BioSpot Core specializes in assays related to the detection, quantification, and qualification of biological “spot” assays and single-cell related analyses, including ELISPOT, colony counting, plaque assays, FluoroSpot, cell viability tests, apoptosis tests, in vivo/in vitro cytotoxicity measurements using cells labeled with fluorescent dyes, histochemistry stains, genotoxicity assays, multi-color intracellular/surface quantification, etc. Immuno/BioSpot Services include: Operator assisted acquisition and analysis Operator unassisted analysis and acquisition (for investigators who have taken the instruction course) Post acquisition data analysis, study, quality control, and file assembly/formatting. ELISPOT / FluoroSpot assay preparation ELISPOT / FluoroSpot assay materials, reagents, and Ab pairs Instrumentation/Software Instruction (permitting unassisted supervised usage of Analyzers). Blank media, disks, and photo printouts ELISPOT / FluoroSpot Assay instruction Free assay consultation Immuno/BioSpot Assay Analysis offered: Quantitative and qualitative ELISPOT assays for cytokines and protein secretions FluoroSpot assays. Fluorescence-based Cell Counting / FluorSpot nuclear counting (to count hyperplasia, tissue infiltration, or cell counting). Microbial assays Colony studies Genotoxic assays Viral plaque assays Stained cell counting from histochemistry slides and sections, including multiple color fluorescent slide images. Assay Descriptions: ELISPOT assays for cytokines and protein production measured at single cell level permitting rapid analysis for exceptional high throughput and extraction of detailed information enumerated from large sample numbers. FluoroSpot assays, permitting directly labeled detection, no enzymatic amplification or substrate, and six-color simultaneous detection permitting fewer cells and reagents. Fluorescence-based Cell Counting: powerful alternative to flow cytometry being: faster (with up to one million cells in an image analyzed in a couple seconds vs. much longer for the same number of cells by flow), more sensitive for rare cells (detection limit one fluorescent cell per million bystander cells i.e. 0.0001%), more economical (every cell is counted, none lost in system), lends itself to high-throughput versus flow, viability counts (live/dead/apoptotic cell counts), FluoroSpot nuclear counting (to count hyperplasia, tissue infiltration, or cells). Microbial assays: microbial load and bioburden testing--enabling colony miniaturization and automated analysis, increasing speed, throughput, and reproducibility. Colony studies permitting automated size-selection and enumeration for clonogenic assays and identifying multipotent progenitor cells. Genotoxic assays for imaging, measuring, and quantifying mutagenic potential by counting cells (mouse lymphoma assays) or microbes (e.g. Ames test). Viral plaque assays performing multiple object-oriented morphometric measurements, enabling user-defined gating by imaging and analyzing the monolayer or bacterial lawn. Quantification of immunohistochemistry and immunofluorescence slides and stains, including multiple-color fluorescent slide images--providing objective quantification along with a recorded image. Consultation and Instruction: Consultation is offered by the Immuno/BioSpot Director, Dr. Christel Uittenbogaart, and Core Manager, Brent Gordon, who can provide protocol workscope planning assistance helpful to investigators who may not have expertise in these assays but wish to incorporate them as a component of their projects. Workshops are offered to investigators for set-up and development of assays and use of acquisition and analysis equipment and software. Quality Control Quality Control for the Immuno/BioSpot core is a particular strength. Possessing the very latest and most sophisticated software, the analysis programs utilize advanced algorithms to assess the entire plate and identify trends in spot intensity, morphology, and size among assigned assay groups, accounting for the variation inherent in distinct experimental groups, yet reducing post-analytic, subjective, user alteration throughout Quality Control. Following analysis, variations inconsistent with plate trends are tagged in Quality Control, requesting verification or signaling the necessity to alter counting parameters to retain valuable or unexpected data. Each alteration from the original counting parameters is recorded in the analysis data file and annotated. All altered counting parameters that fall out of the statistical bounds established in the initial assay analysis are also flagged. The software is compliant with 21 CFR part 11 and GLP regulations. Long term project and analysis trends can also be quantified and observed utilizing the SpotMap software. Finally, the Core Manager has extensive, ongoing experience in assay evaluation and image analysis to help the investigator reduce inaccurate data reporting. OPERATING POLICIES: Once the workscope is determined, the investigator contacts the Core manager, Brent Gordon, who is responsible for all Immuno and BioSpot acquisitions, analyses, materials, and equipment. Brent Gordon can help with study implementation, and schedule time and location to meet the investigator for assay consultation, data acquisition, assay analysis, or material provisions. With two distinct campus locations (Factor 12-541 and 188F BSRB), each featuring different analyzers, accessibility is not a problem. Mr. Gordon may be contacted at both locations from the same phone number and e-mail, and requests 24hr advanced notice to schedule or alter an appointment time to help ensure testing is performed in a timely manner. Dr. Uittenbogaart will assist, as requested, in data interpretation during the project and at the end of the studies.