Projet financé terminé

2005-2008 CEE L’Union Européenne programme FP6-NEST (New and Emerging Science & Technologies)- project AUTOMATION 1,500,000€ (Chief coordinator and principle investigator). “A novel imaging system providing high-content high-throughput multi-dimensional analysis of microscopic biological structure inside non-adherent living cells”
automation
Flow cytometry has the unique advantage that it uses cells in suspension, which makes it an attractive tool to immunologists working on blood cells. Conventional flow cytometry (FC) methods report optical signals integrated from individual cells at throughput rates as high as thousands of cells per second.

This is further combined with the powerful utility to subsequently sort and/or recover the cells of interest. However, these methods cannot extract spatial information. This limitation has prompted efforts by some commercial manufacturers to produce state-of-the-art commercial flow cytometry systems allowing fluorescence images to be recorded by an imaging detector. Nonetheless, there remains an immediate and growing need for technologies facilitating spatial analysis of fluorescent signals from cells maintained in flow suspension.

Since 2002, we have developed di-electric field caging technologies combing micro-fluidic control allowing to manipulate, and trap individual single living cells in suspension. In particular we established the means to use these approaches to make 3D imaging of live cells in suspension, and a joint patent was issued at that time. In 2004 the work was funded as a consortium project in the European Commission, FP6 (framework program) that allowed us to really develop the approach and in particular the mathematics for 3D reconstruction.

The method offers unique possibilities for imaging studies on cells in suspension. In particular, in it’s simplest form uses an approach coined “confocal axial tomography" and uses sequential micro-rotation imaging that yields enhanced isotropic 3-D optical resolution compared with conventional light reconstruction (deconvolution) image data treatment. In it’s more sophisticated form this can be achieved in a continuous micro-rotation tomography mode using any of a variety of more complex, calculation burden intensive reconstruction mathematics developed with our consortium partners.

Today these results are the subject of further development and license negotiations, especially because the methods are relatively rapid and lend themselves to full automation, suggesting utility for 3-D imaging cytometry. This work won the 2005 national prize French Engineers of the Year.