Frédéric LHOMMÉ, Ph. D., ULB
flhomme@ulb.ac.be
Frédéric C. Lhommé was born in 1971 and studied at the Ecole Nationale Supérieure de Chimie et Physique de Bordeaux, France from which he was graduated as Engineer in Chemistry and Physics in 1994. He obtained a PhD in Physics with a specialty in optics from the University of Metz, France in 2000. His research was dealing with the study of the effects of the chromium concentration and crystal composition in LiNbO3:Cr3+ by optical spectroscopy. He did post-doctoral studies about the modelling of the light self –focusing in photorefractive materials at Supélec, Metz, France in 2001 and about the inscription and use of Bragg gratings in optical fibres for sensor applications at the Electromagnetism and Telecommunications Group of the Faculté Polytechnique de Mons, Belgium from 2001 to 2005. Since 2006, he is at the head of the Flow Cytometry unit of the Institute of Medical Immunology of the Université Libre de Bruxelles, Gosselies, Belgium. His research interests are mainly in the development of flow cytometry or related technologies and protocols for research applied to the study of inflammation and transplantation. He published around 25 publications.
Nicolas PASSON, B. Sc.
npasson@ulb.ac.be
Nicolas Passon was born in 1985. He graduated in mathematics and sciences from the high school Saint Joseph at Charleroi. He studied biology during two years at the UMH (University of Mons Hainaut). He obtained a bachelor’s degree in clinical biology in 2008. As a bachelor, he studied the synergy between HDAC inhibitors and Chaetocine in the process of HIV-1 retrotranscription inhibition. He currently works at the CMMI (Center for Microscopic and Molecular Imagery) unit headed by Dr Frédéric Lhommé. He provides technical assistance in informatics, cell culture, molecular biology, flow cytometry, cell sorting and flow cytometry imaging.
Imaging Flow Cytometry
Equipment
The device consists of an Imaging Flow Cytometer ImageStream 100 (Amnis, Seattle) (Figure 1). It allows to acquire several different images per cell and light illumination is provided by 3 laser sources (violet at 405 nm, blue at 488 nm and red at 658 nm) as well as a brightfi eld source.
Figure 1 - ImageStream 100 (Amnis, Seattle)
Figure 2 - Localisation of transcription factor NF-κB in T lymphocyte cells before (A) and after (B) anti-CD3 stimulation. Panels (from left to right), show images recorded with brightfi eld source (BF), NF-κB, nuclear marker Draq5 and the composite view of NF-κB and Draq5.
Description and applications
Imaging Flow Cytometry is a technology which combines in one single device the advantages of the statistical rigour of fl ow cytometry with those of the visual power of microscopy. ImageStream enables fast acquisition of images of cells in a fl ux and allows to analyse them both qualitatively and quantitatively. The analysis is based on criteria such cell morphology, location or intensity of markers on, in or between cells.
The IFC method has been applied to carry out functional studies, phenotyping, cell cycle, apoptosis/nuclear damage, transcription factor nuclear translocation (Figure 2). It has also been usedin a whole range of different disciplines, including immunology, microbiology, parasitology, vaccinology, or cell therapy.
See the poster: POSTER_CMMI_-_Imaging_Flow_Cytometry.pdf
