Multiparametric flow cytometry


 

Flow cytometry is the technique used to quantify the expression of multiple extracellular or intracellular molecules on single cells, allowing the phenotype and functional characterization of complex cell populations. Multiparametric flow cytometry can be particularly suitable for deep analysis of immune responses after vaccination or infection, allowing to measure the frequency, the phenotype and the functional features of antigen-specific cells. The LSR Fortessa ™ X-20 is equipped with 4 lasers (488 nm, 640 nm, 405nm, and 355 nm), allowing to analyze 18 parameters on each single cells.

 

 

 

The high potential of the flow cytometry offers a wide range of applications, including:
– Multi-parametric immunophenotyping of cells
– Assessment of intracellular factors (i.e. cytokines, transcription factors and phospho-proteins)
– Combined assessment of T-cell phenotype and effector function (intracellular cytokine staining in tetramer
– Cell proliferation
– Cell cycle and apoptosis

 

Samples can be analyzed from biological fluids, tissues and cell lines, both of origin human or other species Furthermore, microbiological samples can be analyzed. Data analysis can be performed using specific software (e.g.: FlowJo10, FACSDiva8). It is also possible to perform a computational analysis of multiparametric datasets using software such as FlowSOM, tSNE, Phenograph, in order to characterize all the possible cellular phenotypes in un operator-independent unbiased way.

Ask me for info about this services: Donata Medaglini (donata.medaglini@unisi.it), Annalisa Ciabattini
(annalisa.ciabattini@unisi.it)

 

 

Cell-sorting

The cell sorting is a technique that allows to identify and isolate target cells from the sample. The BD FACSAria™ Fusion instrument allows to isolate up to 4 different cell subpopulations from the initial heterogeneous sample according to the expression of different surface or intracellular molecules. The instrument is equipped with two lasers (488 nm and 633 nm), allowing the simultaneous measurement of 10 different parameters. BD FACSAria™ Fusion is housed inside a Baker Class II biosafety hood which allows to work in sterility and in a safe environment for the operator.
Cell subpopulations can be collected from biological fluids, tissues and cell lines.
Possible applications include:
-Identification and collection of rare sub-populations (i.e antigen-specific lymphocytes)
– sorting of cells for subsequent culture
– single-cell sorting for DNA and RNA measurements
– antigen-specific single B cell isolation

Ask me for info about this services: Donata Medaglini (donata.medaglini@unisi.it), Annalisa Ciabattini
(annalisa.ciabattini@unisi.it)