In our continuing saga of publishing the opinions of leading scientists working in the cutting- edge areas of flow cytometry, we present this month an interview with Dr. Herve Luche, Scientific Director of the Immuno-phenotyping module at the Centre for Immunophenomics – CIPHE (Inserm-CNRS-AMU) in Marseille, France. Dr. Luche and his team are interested in […]
This month’s interview features Dr. John Connolly, Chief Scientific Officer of Tessa Therapeutics, Singapore. Dr. Connolly is a leading immunologist whose research focuses primarily on target discovery for immune modulation. At Tessa Therapeutics, Dr. Connolly and his team are working on virus-specific T-cells (VSTs) and exploring their potential use in human cancer immunotherapy. As Dr. […]
Dr. Adam Laing, PhD joined Professor Hayday’s King’s College London laboratory in 2013 in order to develop the protocols and experiments for the 3i project. Recognised as ambitious and ground-breaking, to date the 3i project has accomplished the immunological phenotyping of 500 knockout mouse lines for the purposes of creating a valuable data set which […]
Dr. Enrico Lugli, ‘Every cell can make a difference’. Dr. Enrico Lugli, PhD is a renowned expert in Flow Cytometry. He is currently principal investigator and head of the Laboratory of Translational Immunology, which is located within the world-famous Humanitas Clinical and Research Centre, Italy. Dr Lugli’s research specialism is the human Immunology System. His […]
An important factor that must be considered for proper panel design is the sensitivity of the reagent. Sensitivity is influenced by the brightness of the fluorochrome and the data spread due to the presence of other antibody conjugates attached to the same cell. Not all fluorochromes have the same sensitivity due to differing brightness. Sensitivity is also impacted by the expression level of the markers being studied. Markers, or antigens, can be classified as primary, secondary or tertiary, depending on their levels of expression. Many candidate antibody-conjugate panels may need to be evaluated to determine which one optimally stains all the antigens under study. This is also important to determine which antibody conjugates reduce the sensitivity. Single stain controls to evaluate and correct for compensation must also be performed. Multicolor experiments can be frustrating but using the right panel can give wonderful results and it is important to invest time and effort into this.
Mahnke, Y.D, Roederer, M, Clin. Lab. Med; 27(3): 469, (2007)