Flow Cytometry (FACS)
The ICGEB Flow Cytometry Facility is supplied with three different instruments, for a wide range of possible flow cytometry applications.
For simple analysis of few parameters, the BD FACSCalibur cell analyser is available, equipped with a single 488 nm argon-ion excitation laser, and three fluorescence reading channels at 530 nm (FL1), 585 nm (FL2), 670 nm (FL3).
More complex multiparametric analysis can be performed on the new BD FACSCelesta cell analyser, equipped with three lasers (violet, 405 nm, blue, 488 nm and red, 633 nm) and the possibility of detecting up to 12 fluorescent channels: 450 nm, 525 nm, 610 nm, 670 nm and 780 nm on the violet laser, 530 nm, 575 nm, 610 nm and 695 nm on the blue laser and 670 nm, 730 nm and 780 nm on the red laser.
Applications of cell sorting can be performed on the BD FACSAria cell sorter, which is also equipped with three lasers (405 nm, 488 nm and 633 nm) and 13 fluorescence reading channels (450 nm, 510 nm, 610 nm, 660 nm, 710 nm, and 780 nm on the violet laser, 530 nm, 585 nm, 616 nm, 695 nm and 780 nm on the blue laser, and 660 nm and 780 nm on the red laser). Sorting of selected cells can be achieved with high efficiency and purity (up to 25,000 events /sec for a 4-way sorting with a purity >98%); in addition, the Automatic Cell Deposition Unit allows sorting on slides and plates (6, 12, 24, 48, 96 and 384 wells plates are accepted).
Applications of flow cytometry include, among others, immunophenotyping of cell populations for expression of specific markers, studies on gene expression, analysis of cell viability and proliferation, analysis of cell cycle by means of DNA-specific dyes which allow quantitation of cellular DNA content, studies of ion fluxes in response to specific stimuli and many more; selected sub-populations can be sorted for analysis or sub-cloning or co-cultivation of selected populations.
A rich library of manuals and application procedures can be found in the BD Biosciences site:
For further information:
Marco Bestagno, Cellular Immunology Laboratory