accumulative difference image protocol for particle tracking in fluorescence microscopy tested in mouse lymphonodes累积差分图像在荧光显微镜测试协议为粒子跟踪鼠标lymphonodes.pdfVIP

Accumulative Difference Image Protocol for Particle Tracking in Fluorescence Microscopy Tested in Mouse Lymphonodes 1 1 1 1 1 2 Carlo E. Villa , Michele Caccia , Laura Sironi , Laura D’Alfonso , Maddalena Collini , Ilaria Rivolta , 2 3 3 3 1 Giuseppe Miserocchi , Tatiana Gorletta , Ivan Zanoni , Francesca Granucci , Giuseppe Chirico * ` ` 1 Dipartimento di Fisica, Universita degli Studi di Milano Bicocca, Milano, Italy, 2 Dipartimento di Medicina Sperimentale, Universita degli Studi di Milano Bicocca, Monza, ` Italy, 3 Dipartimento di Biotecnologie e Bioscienze, Universita degli Studi di Milano Bicocca, Milano, Italy Abstract The basic research in cell biology and in medical sciences makes large use of imaging tools mainly based on confocal fluorescence and, more recently, on non-linear excitation microscopy. Substantially the aim is the recognition of selected targets in the image and their tracking in time. We have developed a particle tracking algorithm optimized for low signal/ noise images with a minimum set of requirements on the target size and with no a priori knowledge of the type of motion. The image segmentation, based on a combination of size sensitive filters, does not rely on edge detection and is tailored for targets acquired at low resolution as in most of the in-vivo studies. The particle tracking is performed by building, from a stack of Accumulative Difference Images, a single 2D image in whi