Fluorescence Confocal Microscopy imaging denoising with photobleaching.
The Fluorescence Confocal Microscopy (FCM) is nowadays one of the most important tools in biomedical and pharmaceutic research. The main advantage of this technique over the traditional bright field optical microscopy is the fact that it allows the selection of a thin cross-section of the sample by rejecting the visual information coming from the out-of-focus planes. However, the small amount of energy radiated by the fluorophore and the huge light amplification performed by the photon detector to capture this visual information introduces a type of multiplicative noise described by a Poisson distribution. Additionally, the radiation efficiency of the fluorophore decreases with the time, an effect called photobleaching, leading to a decrease in the image intensity along the time. In this paper, a reconstruction algorithm is proposed where the multiplicative noise and the photobleaching effect are modeled. The goal is to obtain the morphology and the intensity decay rate across the cell nucleus from a sequence of FCM images. The reconstruction algorithm is formulated as an optimization problem where a convex energy function is minimized. Tests using synthetic and real data are presented to illustrate the application of the algorithm and the effectiveness of the results.
Rodrigues
Isabel
I
Instituto Superior de Engenharia de Lisboa, Portugal. irodrigues@isr.ist.utl.pt
Xavier
Joao
J
Sanches
Joao
J
D000465:Algorithms; D005456:Fluorescent Dyes / Q000528:radiation effects; D007089:Image Enhancement / Q000379:methods; D007090:Image Interpretation, Computer-Assisted / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008962:Models, Theoretical; D038761:Photobleaching
D005456:Fluorescent Dyes