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|  Xavier University of Louisiana |
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| Xavier University of Louisiana |
The Cell Analysis & Immunology Core Facility is a customer oriented service dedicated to supporting the research needs of the investigators of the Louisiana Cancer Research Consortium (LCRC). It provides state-of-art instrumentation and expertise for addressing experimental issues related to single cells, nuclei and chromosomes.
In addition to instrumentation, the Facility intends to collaborate with the investigators and contribute to all aspects of the research process. This includes consultation in experimental design, technical assistance in the operation of instruments, trouble shooting, data analysis and storage, interpretation of results and preparation and production of presentation graphics. The Facility is also communicating with investigators in other institutions and is devoted to introducing and developing new applications on our flow cytometers. In this way, investigators are provided with unlimited possibilities to answer questions regarding nature and function of cells by flow cytometry.
Member:
Finkel-Jimenez, Beatriz, Ph.D.
Role:
Technical Director (LSU), Cell Analysis & Immunology Core
Member:
Ochoa, Augusto, M.D.
Role:
Scientific Director, Cell Analysis & Immunology Core
Member:
Price, Mary, B.S.
Role:
Technical Director (Tulane), Cell Analysis & Immunology Core
"It is a combination of classic AID reader unit and the AID EliSpot software to acquire, analyze, and quantify the spots according to the user's settings."
Available at LSU site.
"The newest cell sorter which is equipped with three three solid state lasers. The computer is capable of collecting 11 parameters simultaneously (2 scatter signals and 9 fluorescent parameters). It is designed to be a high precision sorter, which allows for acquisition rates of up to 70.000 cells per second and sorting rates up to 40,000 cells per second while maintaining high purity and cell recovery yields. Cells can be collected under non-aseptic or aseptic condition. An automated cell delivery system (ACDU) permits sorting individual cells for direct cloning into 96 well plates."
Available at the Tulane site.
"A compact system rapidly detects and quantifies concentrations of secreted proteins, proteins in cell lysates, and cell-associated proteins using small sample volumes. It can be used to detect apoptosis, cytokine and chemokine profiling and phosphorylation of key signal transduction proteins by multiplexed BD cytometric bead array (CBA) products. FACSArray is a very flexible system suitable for many different types of application and this system is fast and requires minimal training."
Available at the Tulane site.
"The BD FACSCalibur system combines unique dual-laser technology. Designed specifically to support a wide range of applications, the BD FACSCalibur system is a fully integrated multiparameter system that offers investigators the performance and flexibility required for a variety of research applications."
Available at LSU site.
"A three-laser bench-top flow cytometer designed with all-digital signal collection and processing and a fixed alignment fiber optical system engineered to achieve the greatest amount of signal detection for each laser. The standard optical filters on the LSR allow the detection of (but are not limited to) the following fluorophores: PE-Cy7, PE-Cy5.5, PE-TexasRed, PerCP, PerCP-Cy5.5 PE, FITC, PI, APC-Cy7, APC, Hoechst, DAPI, Cascade Blue, Pacific Blue, Alexa Fluor 405 and Alexa Fluor 430. It can simultaneously detect 10 parameters (2 scatter and 8 fluorochromes). Additionally, the LSU and Tulane sites both have an HTP attachment which allows high through put analysis of 96 well plates."
Available at both the Tulane & LSU sites.
"This is a fully integrated system combining hardware, Bio-Plex Manager software, system validation and calibration tools, assays, and beads. The system ensures accurate and reproducible results."
Available at the LSU site.
"The auto MACS separator is a computer controlled magnetic cell sorter for isolation of virtually any cell type. The system is compatible with all MACS cell separation regents. There are several separation programs available allowing different cell selection strategies. Employing the MACS magnetic cell separation technology the auto MACS separator is capable of sorting more than 10 million cells per second from samples of up to 109 total cells."
Available at LSU site.
"The Luminoscan Ascent is a sensitive and flexible microplate luminometer. Combined with the powerful Ascent software, it handles both glow and flash type reactions. It offers great advantages for different types of assays. These include reporter gene assays, immunoassays with luminescent substrates, cytotoxicity and cell proliferation assays, Intracellular Ca+2 assays, ATP assays, DNA quantification."
Available at LSU site.
"The facility adheres to a file saving structure based on the investigators email/logon ID. Each user will obtain a logon ID and a password (usually their last name) for access to the instruments. The investigators need to create a folder with his last name. This is where the data will be saved. The FCS files will be named as mmddyyxxxF, where m=month, d=day, y=year, xxx=users' last name, F= name for instrument used in the experiment. Data is backed up at the end of each day to an archival computer. At the Tulane site, investigators can access this computer over the network to retrieve data for approximately 6 months. At LSU investigators will need a flash drive or CD to retrieve data, as no network access is available. All data is permanently archived to CD-ROM storage disks."
"The sorter BD FACSAria is run exclusively by the operator and highly trained authorized users. The fluidic systems are cleaned and sterilized with antiseptic detergent to maintain aseptic conditions and eliminate crossover contamination between samples."
"An operator is available for running samples on BD LSRII and FACS Array. The facility can also assist investigators with the setup and running of samples. Investigators are encouraged to learn how to run their experiments on the LSRII and FACS Array at the Tulane site to enable independent usage of the facility allowing for the flexibility to schedule time for analysis during the evenings and weekends. For this purpose, users need to be trained and demonstrate that they know the proper set up, running and shut down procedures. At the LSU site, only trained operators can run the equipment, therefore, arrangements for services need to be scheduled.
Among the available analyses:
1. immunophenotyping (cell surface antigens and receptors)
2. intracellular proteins, cytokines, calcium flux, pH and free fatty acid,
3. signal transduction pathways,
4. DNA and RNA content, cell cycle, DNA synthesis,
5. apoptosis (programmed cell death) analysis, based on:
- mitochondrial membrane potential
- down regulation of BCL-2
- activation of caspases (cysteine proteases)
- cleavage of the DNA repair enzyme PARP
- DNA fragmentation (TUNEL Assay with anti-BrdU labeling)
- translocation of phosphatidylserine from the inner cytoplasmic membrane to the cell surface (Annexin-FITC/PI)
The facility will assist with the retrieval and analysis of data and interpretation of results. There are several packages of software available to analyze list data, including Cell Quest, FlowJo, Microsoft Word, Excel, PowerPoint, as well as Photoshop."
Facility provides training for instruments, data analysis, and specific applications.
"The same flexible, yet easy to use Thermo Scientific Ascent Software is used to control all members of the Thermo Scientific Ascent family of microplate readers."
"BD CellQuest Pro software allows you to acquire and analyze data from your flow
cytometer on a Macintosh® computer. In BD CellQuest Pro software, an Experiment document contains elements such as plots, regions, gates, statistics, markers, and annotated text. Data files are not saved in the Experiment document. The software saves a reference to the data files that contain the information needed to recreate the plots and stats on the Experiment document."
Flow cytometry data and statistical analysis software.