Flow cytometry could be defined as the measure of cells in fluidics, that means it could be possible to distinguish a cell in constant fluidics. The Flow cytometer is an instrument that allows measure three components in one simple cell: 1) the relative size 2) the internal complexity and 3) the expression of proteins of interest through antibodies coupled to fluorophores and another fluorescent proteins . The equipment is composed by Optical system, relative to the use of laser at different wavelengths which excites the fluorophores in the sample and a collection of different detectors. Fludics, this component is one of the more important in flow cytometry, because is necessary to the sample being in a constant fluidic allowing the alignment of sample components and each component cross through the laser individually. The last component is the electronic component, which is capable to transform the analog signal receive from the sample excitation to a digital form to be visualized and processed in a computer. In our service we count with three flow cytometers and one cell sorter.
FACSCalibur: One of the most basics instruments that we have in the unity, this equipment is capable to analyze 4 colors (FL-1, FL-2, FL-3 y FL4) simultaneasly additionally to side scatter (SSC) and forward scatter (FSC). The configuration of this equipment allows the measurement of each component in height (H). This cytometer has the next filters: FL1 530/30 (FITC), Fl2 585/42 (PE), FL3 670 (PercP) excited by blue laser (488nm) and FL4 661/16 excited by red laser (635nm). The resolution of this cytometer is enough to separate clearly monocytes, lymphocytes, granulocytes and detect bacteria.
FACSCanto II: Is the most used cytometer inside of the unite, this equipment is capable to analyze 6 different colors simultaneously additionally to side scatter (SSC) and forward scatter (FSC). This cytometer has the configuration to capture all measures in the three components, height (H), area (A) and width (W), which simplify the singlet cell separation. This cytometer has four detectors for the blue laser: 530/30 (FITC), 585/42 (PE), 670 (PercP, PercP-Cy5.5), 780/60 (Pecy7) and 2 detectors for the red laser (633nm) 660/20 (APC) y 780/60 (APC Cy7). The resolution is FSC 1.0mm and SSC 0.5mm.
LSRFortessa X20: The newest cytometer that we have in the platform. This cytometer is capable to analyze 18 colors simultaneously, additionally to side scatter (SSC) and forward scatter (FSC). The configuration of this equipment allows to make measurement of the all parameter in the three principal components: height (H), area (A) and width (W) which make simple the separation of singlet cells. This cytometer has: 6 detectors for the blue laser (488nm), 525/50 (FITC), 575/25 (PE), 610/20 (Pe-CF594), 670/30 (PeCy5), 695/40 (PercP, PercP-Cy5.5) y 780/60 (Pecy7). Four detectors for the red laser (640nm) 670/30 (APC), 685/35 (APC Alexa 680), 730/45 (APC Alexa 700) y 780/60 (APC Cy7). Six detectors for the violet laser (405nm) 450/50 (BV421), 525/50 (BV510), 610/20 (BV605), 670/30 (BV650), 710/50 (BV711) y 780/60 (BV786) and 2 detectors for the ultraviolet laser (355nm) 379/28 (BUV395) y 740/35 (BUV737). The resolution of SSC is 0.5mm.
FACSAria II: This cytometer besides of have all the capacity of every cytometer has the capacity to separate specifics population of interest. This cell sorter has the capacity to separate 4 populations simultaneously. The cytometer configuration allows the measure of the height (H), Area (A) and width (W) component for each parameter. This cell sorter has 5 detectors for the blue laser (488nm), 530/30 (FITC), 585/42 (PE), 616/23 (Pe-CF594), 695/40 (PercP, PercP-Cy5.5) y 780/60 (Pecy7). Two detectors for the red laser (640nm) 660/20 y 780/60 (APC Cy7) and 2 detectors for near Ultraviolet laser (375nm) 450/40 (DAPI) y 530/30 (Pacific Orange). The resolution of the cytometer is FSC 0.5mm and SSC 0.5mm.