Three-dimensional reconstruction of fluorescent lymphocyte cells

The complete process, image acquisition, deblurring, 3D reconstruction and analysis has been tested on 160 normal T lymphoid cells marked with monoclonal antibodies CD4-CD8. The images are acquired from a conventional optical microscope illuminated by a mercury vapor lamp. A set of 16 serial image cuts is first grabbed by a high sensitivity Silicon Intensified Target (SIT) camera fitted on the microscope. The 2D image cuts are then deblurred using an original deconvolution method developed in the laboratory. The 3D reconstruction is processed slice by slice using a variant of the 2D Delaunay triangulation. To get a correct triangulation in the case of our specific application the following stringent requirements must be met. First, we exclude the possibility of joining two contours if we meet an empty cut between them, and secondly we must avoid branching points and finally, we state that reconstructed shapes may not have holes. The 3D reconstruction highlights some qualitative characteristics of the cells. So we can show that the fluorescent spots are either scattered on the membrane surface and present a regular radial arrangement or on the contrary are confined in a restricted area. Accordingly we propose to introduce a spatial coefficient in order to characterize such a volume distribution. In a further development we plan to obtain discriminant descriptors of the immunological status of hematopoietic cells.© (1995) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.