BD BioSciences, Belgium
FACS Canto™ II system
Fluorescence Detector Design: Reflective optics with single transmission filter in front of each PMT
Blue Laser Dyes: FITC, PE, PerCP or PerCP-Cy™5.5, PE-Cy™7 (525, 575, 678 or 695, 785 nm)
Red Laser Dyes: APC, APC-Cy7 (660, 785 nm)
Sample Injection: Direct into flow cell
Max Particle Size 50 μm
Sample Flow Rate, Min 10 μL/min
Sample Flow Rate, Max 120 μL/min
Sample Acquisition Rate: 10,000 events/second, 6 compensated fluorescence parameters and 2 scatter parameters
Sample Dead Volume: 30 μL (BD Falcon™ tubes 12 x 75-mm)
Integrated fluidics cart and compressor
Provision of fixed optical alignment procedure
The BD FACS™ Sample Prep Assistant II
TCell components are fluorescently labelled and then excited by the laser to emit light at varying wavelengths. The fluorescence can then be measured to determine the amount and type of cells present in a sample.
Cell counting, cell sorting, biomarker detection
Users can follow either of the standardize protocols as per the requirement of the user i.e. Direct staining, Indirect staining, intracellular staining or cell-cycle staining.
Direct staining: Live or fixed cells are incubated with directly labeled antibodies against cell surface antigens.
Indirect staining: If there is not a directly labeled antibody available, or user wish to amplify their signal he/she can do indirect staining. This is where user stain a cell with a primary antibody against the antigen of interest and visualize using a labeled secondary antibody which recognizes the primary.
Intracellular staining: In order to detect antigen not present on the cell surface, cells have to be fixed and permeabilized to disrupt the cell membrane and allow entry of the antibody. Antigens can be then directly or indirectly labeled. Various methods are optimal depending on the antigen and antibody used.
Cell-cycle staining: Measuring DNA content for cell cycle analysis requires fixation and permeabilization of the nuclear membrane.
Users can also follow the procedure for cell preparation, cell activation to determine immune competence, marker upregulation, cytokine release and proliferation by flow cytometry.