Flow Cytometer

Suitable for single cell analysis. Equipped with acusting focusing, excitation laser at 488 nm, detection filters to detect FSC, SSC and fluorescence emission for green, yellow, red and far red. Implementable with more detection filters and excitation lasers. Analysis at flow rate between 12,5 µL/min and 1.000 µL/min, up to 35.000 events/second. 80 MESF for FITC, ≤30 MESF for PE, ≤70 MESF for APC.
https://www.thermofisher.com/it/en/home/life-science/cell-analysis/flow-cytometry/flow-cytometers/attune-nxt-flow-cytometer.html

The BD FACSCalibur Flow Cytometer allows the measurement off cell characteristics, which can include cell size, cell count,cell cycle and name. The most critical requirement for efficient and effective flow cytometry analysis is that the sample be a single-cell suspension. This helps ensure that everey cell is analyzed independently. The suspension of cells or particles is aspirated into a flow cell where, surrounded by a marron fluid stream, they pass one at time through a focused laser beam. The light is either scattered or absorbed when in strikes a cell. Absorbed light of the appropriate wavelenght may be re-emitted as fluorescence if the cell contains a naturally fluorescent substance or one or more fluorochrome-labeled antibodies are attached to surface. Various immunofluorescent dyes or antibodies can be attached to the antigen or protein of interest. This approach makes flow cytometry a powerful tool for detailed analysis of complex populations in a short period of time.

This platform at CLNS-IIT@Sapienza provides basic support to the projects in the field of flow
cytometry, as well as help in the development of novel protocols and methodology. The laboratory
technology has the main purpose to characterize and isolate a variety of cell population both of human
and mouse origin.
Flow Cytometry platform setup
The flow cytometry laboratory is equipped with two laser-based instruments which technology allows
simultaneous multiparametric analysis of thousands of cells for second for rapid analysis and/or
isolation of complex cell populations. This is achieved by the use of different labeled antibodies or
dyes to target specific molecules express at the surface or in the cytoplasm of the cell, giving a
screening able to describe in detail a complex sample or to identify low frequency subset of cells.
Through these instruments, a variety of fluorochromes attached to different antibodies can screen the
molecular components of a cell first to characterize it (flow cytometer) and then to specifically select
for isolation (cell sorter). The following machinery have been purchased:
• The BD LSRFortessa cell analyzer is a 5 laser cytometer (488nm, UV 355nm, V 405nm,
561nm and 640nm) and 18 fluorescence detectors running a BD FACSDiva software on XP.
The main feature of the cell analyzer is to characterize a given cell population.
• The BD FACSAriaIII is a 4 laser cell sorter (488nm, Near UV 375nm, 561 nm and 640nm)
and 10 fluorescence detectors running a BD FACSDiva software on XP. The main feature of
the cell sorter is to isolate a specific population.