1. Advice to users about flow cytometry service.

  • Initiation and training available for users that use the equipment of the service for the first time.
  • Supervision of the daily operations in analyzer flow cytometers.
  • Support to the users on possible problems with equipment.
  • Support to the users about methodology and experimental design.

2. Applications

  • Complex multiparameter analysis: immunophenotyping by identification of surface antigen is possible by combining fluorochromes compatible with laser lines and filters. Immunophenotyping refers to the use of immunologic tools, such as fluorochrome-coupled antibodies, for the specific detection of antigens located into and on the surface of the cell. It enables to identify, list, and characterize any individual cell or cellular subcomponent. It has very important applications such as the identification of hematologic diseases, hematologic disorders, transplantations, diagnosis of infectious diseases, detection of fetomaternal hemorrhage, allergy testing based on the activation of basophils, platelet activation, diagnosis and monitoring of autoimmune diseases, identification and quantification of autoantibodies, identification of multidrug-resistant phenotypes in cancer, quantification of soluble proteins,…
  • Reporter genes detection: One of the most important techniques in the study of a gene of interest’s expression is the reporter gene fusion, which allows a rapid and easy determination of the activity of a promoter. Green fluorescent protein (GFP) is frequently used as a reporter gene in cytometry.
  • Study of cellular death process: It enables to differentiate whether the cell is in the early or late stage of apoptosis, or conversely whether cell death occurred by necrosis. For that end, it is quantified the amount of Annexin-V binding phosphatidyserine of the surface of apoptotic cells, or the internalisation of propidium iodine (PI) or 7-Aminoactinomycin D (7-AAD), due to the formation of pore in the cell membrane, which is characteristic of the necrotic cells.
  • Cell cycle and cell proliferation: The studies on cell cycle show several cell division phases, according to the amount of DNA, which allows to calculate indices and ratios in the different peaks. Flow cytometry is able to distinguish the different phases of the cell cycle; G0/G1 phase: the cell is in quiescent stage and preparing for cell division with a DNA content of 2n; S-phase: is the phase of DNA synthesis; and the G2/M phase: during which the amount of DNA in the cell is doubled (4n). The IP, Hoechst, 7-AAD, and BrdU labeling are usually the most used, among others.
  • Functional assays: Cytometry allows the quantification of the dynamic properties of cells that can be rapidly modified thanks to fluorescent sensors, whose optical and distribution properties change in response to the phenomenon of interests. For example, ROS detection, or fluo/indo ratio-metric measurement to study calcium flux.

3. Cell Separation.

Separation of cells or homogeneous cell population are performed for subsequent biochemical, molecular, or cell differentiation assays of populations of interests.