Flow Cytometry (FACS)
The ICGEB Flow Cytometry Facility offers support to perform a range of flow cytometry applications.
Simple analysis of few parameters are performed using BD FACSCalibur, 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).
For multiparametric analysis we use a BD FACSCelesta cell analyser, equipped with three lasers (violet, 405 nm, blue, 488 nm and red, 633 nm) and the possibility to detect up to 12 fluorescent channels:
Cell sorting is performed on the BD FACSAria cell sorter, equipped with three lasers (405 nm, 488 nm and 633 nm) and the possibility to detect up to 13 fluorescence channels.
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).
We offer initial training and support to design panels to identify and isolate rare population out of heterogeneous samples, with a specific focus on the identification of subpopulation of immune cells infiltrating tumors (link to cytofluocourse).
We established efficient sterile sorting of CRISPER/CAS9 genome edited cells for subsequent expansion and subcloning. Other possible applications include analysis of gene expression, cell viability and proliferation, cell cycle, ion fluxes in response to specific stimuli.
We provide advices and dedicated training sessions in the use of flow cytometry, instrument operation and post-acquisition data analysis on the Flow-Jo software.