CARC Biospecimen Processing & Analysis Laboratories
The CARC Biospecimen Processing & Analysis (BSPA) Core has 3100 sq. ft. of modern laboratory space on the 3rd floor of the Clinical Sciences Research Building (CSRB) of the LSU Health Sciences Center in New Orleans, where experiments and analyses proposed in the different Research Components and Pilot Projects are performed. The BSPA Core is a multidisciplinary Core that consists of a Flow cytometry, Molecular Biology/Analytical and Cell Culture Laboratories dedicated to support SIV/HIV/Alcohol research. The laboratories are fully equipped with fume hoods, air, gas and vacuum lines. Water is purified centrally and run through Millipore ion exchange and activated carbon beds in the laboratories. Additional laboratory space (approximately 2800 sq. ft. at LSUHSC) holds additional equipment that will be used by the proposed studies and shared with other faculty members.
Flow Cytometry Laboratory
Technical Director: Constance Porretta, BS, I(ASCP)
The Flow Cytometry Laboratory is instrumented with 3 cell sorters and 2 analysis cytometers with high throughput sampling capacity. Our cell sorters have a maximum of 15 fluorophore detection capacity, and the analyzers have a maximum of 18 fluorophore detection capacity. We routinely perform polychromatic cell analysis and sorting, immunophenotyping, intracellular and functional markers, cytometric bead array assays, DNA ploidy, cell cycle, and proliferation analysis. Our laboratory offers in-house training and remote access to analysis programs such as FlowJo, Diva, FCAP, and Modfit.
- Antigen-specific T cell phenotyping, cytokine production, and SPICE analysis.
- CFSE analysis of proliferating cells over 10 cell divisions.
Molecular Biology/Analytical Laboratory
Technical Director: Jane A. Schexnayder, BS
The Molecular Biology/Analytical Laboratory is a state-of-the-art facility capable of performing most major molecular biology techniques. Examples of our capabilities include gene cloning, RT-qPCR and standard curve generation, Western blot analysis, ELISA, ELISPOT, Multiplex protein arrays, and DNA methylation activity assays. Development of mRNA semi-quantification using Multiplex technology on fixed, permeablized flow-sorted cells is among our current innovation.
- RT-qPCR standard curve amplification plots for SIVgag.
- Plotted standard curve of SIVgag has 100.7% efficiency and an R2 value of 0.999
- Western blot analysis of phosphorylated p42/44 MAP kinase. (1) Marker, (2) Saline ip/D5W it, (3) Saline ip/G-CSF it, (4) Alcohol ip/ G-CSF it, (5) Saline ip/D5W it, (6) Saline ip/G-CSF it, (7) Alcohol ip/ G-CSF it, (8) Saline ip/D5W it, (9) Saline ip/G-CSF it, (10) Alcohol ip/ G-CSF it
- Mutiplex standard curve for IL-6 analysis.
Cell Culture Laboratory
Technical Director: Rhonda Martinez, BS
The Cell Culture Laboratory is a specialized BSL-2 facility equipped to handle samples obtained from both human tissues and rhesus macaques. Additional laboratory space is dedicated to the culture and analysis of cell lines and primary cells from other non-primate model organisms. Techniques commonly used in the laboratory include in vitro PBMC stimulation for downstream assays, isolation of primary bone marrow mesenchymal stromal cells from rhesus macaques, in vitro SIV/HIV infectivity assays, and time-lapse microscopy of living in vitro cultures in a humidified and heated stage microscope. We are currently developing lentiviral shRNA delivery systems to prepare customized knockdown capability to CARC investigators.