V Congreso Latinoamericano de Ingeniería Biomédica (CLAIB2011)

Evaluation of the Expression Level of a Fluorescent Protein in Single Cells through Digital Image Processing
Selvaggio Gianluca, Ceroni Francesca, Giordano Emanuele, Lorenzo Ginori Juan Valentín

##manager.scheduler.building##: Palacio de Convenciones de La Habana
##manager.scheduler.room##: Sala 8
Fecha: 2011-05-20 09:00  – 09:15
Última modificación: 2011-04-28 01:47


Fluorescent signals are widely used in life sciences to investigate cellular behaviors. Using a fluorometer is a reliable approach to measure the amount of a specific fluorescent radiation emitted by a cellular population. On the other hand, measuring fluorescent signals at the single-cell level requires the use of properly equipped microscopes to acquire images for further analysis. Here we present a digital image processing (DIP) approach to analyze these images. More in detail, a DIP system was developed to analyze photographic images of fluorescent Escherichia coli expressing a Green Fluorescent Protein (GFP). This system includes stages of noise filtering, uneven illumination correction, segmentation, detection of cell aggregates and measurement of the level of fluorescence of the selected cells. To accomplish this task, well established DIP algorithms for the different functions were adapted to the specific conditions of the experiment under study. In the results section, images are presented that reveal the behavior of the algorithms employed, as well as the calculated distribution of single cell fluorescence. In regard of the adequate selection of valid cells (i.e. deletion of aggregates), the system effectiveness was evaluated through comparison to the results obtained by human analysis, and expressed quantitatively. An assessment on the perspectives of further improvements to the developed system is finally proposed.