Flow Cytometry Unit

Researcher:  Despina Smirlis, PhD, researcher C’
Email:  penny@pasteur.gr, Tel: +30 210 6478841

Research Assistant: Evangelia Xingi, PhD
Email: exingi@pasteur.gr, Tel: +30 210 6478834

Overview

Overview

 

The Flow Cytometry facility of the Hellenic Pasteur Institute (HPI) is equipped with a FACS ARIA cell sorter and a FACS Calibur flow cytometer. The facility is supported by the junior researcher Despina Smirlis specialising in the field of Molecular Microbiology and the Research Assistant Dr. Evangelia Xingi. Aim of the facility is to provide advanced flow cytometry and sorting facilities to the HPI scientists as well as to visiting scientists.Flow cytometry is an automated, quantitative and dynamic and multi-parameter way of analysing the physical and chemical properties of particles and cells, based on light scattering by the particles or fluorescence emmission. A flow cytometer is characterized as a successful combination of an haematological analyzer and a fluorescence microscope, that uses the last microscopy field achievements and biochemical analysis as well as computer evolution.Hydrodynamic focusing of the sample through the flow chamber
 

Simplified illustration of flow cytometry

Equipment

Equipment/ Infrastructure

 

Current equipment in the Flow Cytometry facility includes:

1. BD-Biosciences FACS Aria flow cytometer/cell sorter

  The facility is equipped with the BD FACSAria™ flow cytometer/cell sorter that sets a new standard for high performance flow cytometry. Based on a revolutionary new design in instrumentation, this easy-to-use benchtop system delivers high-speed cell sorting and multicolor analysis.The HPI FACSAria is equipped with an octagon containing six PMTs, which detect light from the 488-nm (blue) laser, and a trigon containing two PMTs, which detect light from the 633-nm (red) laser. The flow cytometer can detect up to 7 fluorescent parameters and two scatter parameters at a time. The sorted samples are collected into up to four 5ml tubes.
BD FACS Diva software 6 is used for data acquisition and analysis that can also be operated by a second PC workstation.

2. BD-Biosciences FACSCalibur flow cytometer The HPI FACSCalibur flow cytometer is equipped with an Argon laser 488 nm and a red diode laser 635 nm. The flow cytometer is connected to a BD FACStation™ v6.0x for Mac OS® X, which uses the BD CellQuest™ Pro software for data acquisition and analysis. The system is also equipped with the ModFitLT software for cell cycle analysis.

Table of instrument configurations

Regulations

Regulations

 

  • HPI users

The use of all the equipment in the Flow Cytometry facility is free of charge for HPI scientists.

FACS Aria cell sorter: sorting services are provided only by the facility Research Assistant, so all users should get in contact with the responsible scientists.

FACS Calibur flow cytometer: All users should fill the online booking file.

 

  • External users

The facility equipment can be used by external users, after communication with the responsible scientists for reservation of the instruments.
There is a charge of 20 € / hr for the use of either of the two systems.