FITC-labeled heparin binding to live, apoptotic, and necrotic Jurkat cells and to nuclei prepared from Jurkat cells as determined by fluorescence microscopy.

Cell death was induced by Camptothecin, and cells were separated into live, apoptotic, and necrotic populations by cell sorting. The upper line of micrographs [1] shows typical examples of the binding of FITC-LMWH to each population, as observed by confocal microscopy. [1A] Live cells, [1B] apoptotic cells, [1C] unlysed necrotic cells, and [1D-E] lysed necrotic cells. FITC-LMWH is seen as green fluorescence, and the red fluorescence resulted from binding of PE–annexin V and 7-AAD. The middle line of micrographs [2] shows the binding of FITC-LMWH as determined by fluorescence microscopy. DAPI, (blue fluorescence); FITC-heparin (green). [2A] Live cells, [2B] apoptotic cells, [2C] unlysed necrotic cells, and [2D-E] lysed necrotic cells. The lower line of micrographs [3] shows FITC-heparin binding to purified nuclei prepared at timed intervals after the addition of Camptothecin. Nuclei were stained with FITC-heparin (green) and DAPI (blue). [3A] Zero time, [3B] 30 minutes, [3C] 1 hour, [3D] 2 hours, [3E] 4 hours, and [3F] 6 hours.