​​​​​​​Stellenbosch University Immunology Research Group​​

​Platforms​

Flow Cytometry is a laser based technology used to analyse the physical and chemical characteristics of cells or particles. Our highly trained staff offers a wide range of expertise in:

• Study and panel design as well as discussing gating strategies for multi-colour experiments.
• Processing of different sample types such as whole blood cells, PBMCs, cultured cells and cell lines, pericardial fluid, bacteria.
• Acquiring of samples using different Flow Cytometers:
• BD FACSCanto II: 3 laser (405nm violet laser, 488nm blue laser, 633nm red laser), 8 colours
• BD LSR II: 3 laser (405nm violet laser, 488nm blue laser, 633nm red laser), 12 colours
• BD FACSAria IIu cell sorter: 3 laser (405nm violet laser, 488nm blue laser, 633nm red laser), 8 colours
  
• BD FACSJazz cell sorter: 3 laser (405nm violet laser, 488nm blue laser, 561nm yellow/green laser), 6 colours
• Amnis ImageStreamX: 8 laser (375nm, 405nm, 488nm, 561nm, 592nm, 642nm, 730nm, 785nm), 12 channels
  
• Analysis of samples using FlowJo.

Our staff is experienced in the following applications:

• Immunophenotyping of whole blood or cells for innate and adaptive cell frequencies and subsets, including maturation, trafficking, activation, proliferation and chemokine and cytokine expression.
• Determining the percentages and absolute counts of T, B, and natural killer (NK) cells as well as the CD4 and CD8 subpopulations of T cells in peripheral blood using BD Multitest™ 6-color TBNK reagent and BD Trucount™ tubes.
• Live/ dead discrimination of bacteria.
• Infectious and non-infectious cell sorting of cells and bacteria.

If you consider using Flow cytometry and need guidance in setting up your experiments, panel design, acquiring and analysis of your samples; contact our Flow Cytometry expert Andrea Gutschmidt.

Luminex is a dual-laser flow-based detection technology and allows for analysis of up to 50 analytes in a single well.

Our staff has vast experienced in polystyrene or paramagnetic beads assays using the Bio-Plex 200 and Bio-Plex MAGPIX instruments.

If you wonder if Luminex is the right platform for your assays, and if you strive for accuracy and reproducibility speak to our Luminex expert Dr Novel Chegou.

The Agilent Seahorse Extracellular Flux (XF) Analyzer is the world's most powerful metabolic analyzer. It is based on measuring rates of change in pH and Oxygen concentration in the two metabolic pathways by which cells use to generate energy, i.e. Glycolysis and Oxidative Phosphorylation respectively.

It offers the opportunity for functional metabolic analysis essential to tell the full story of cellular processes and pathologies in real time within minutes. The Seahorse technology enables the measurement of metabolic function and energy phenotype in patients-derived, and other precious samples.

We run samples in our lab using the Agilent Seahorse XFp Analyzer. For clinical appraisal of cellular energy metabolism, the following questions can be answered using the Seahorse analyzer:

• XF Cell Energy Phenotype Test, measure energy phenotype and possible metabolic switches:
• Define XF Metabolic Phenotypes (Glycolysis or Mitochondrial Respiration)
• Measure XF Metabolic Potentials (Cells ability to meet up with its energy demands)
• Study Metabolic Switches (From Glycolysis to Oxidative phosphorylation and vice versa)
• XF Glycolysis Stress Test, measure the key parameters of glycolytic function:
• Glycolysis
• Non-glycolytic Acidification
• Glycolytic Capacity
• Glycolytic Reserve
• XF Mitochondrial Fuel Flux Test, measure the key parameters of mitochondrial fuel use:
• Fuel Dependency
• Fuel Flexibility
• Fuel Capacity
• XF Cell Mitochondrial Stress Test, measure the key parameters of mitochondrial function:
• Basal Respiration
• ATP Production
• Maximal Respiration and Spare Capacity
• Proton Leak
• Study metabolic fluxes of cells treated with different drugs through injection ports

If you consider using the Seahorse technology for metabolic analysis, contact our specialist Dr Nelita du Plessis.