title: Accumulative difference image protocol for particle tracking in fluorescence microscopy tested in mouse lymphonodes.
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 which the motion of the whole set of the particles is coded in time by a color level. This algorithm, tested here on solid-lipid nanoparticles diffusing within cells and on lymphocytes diffusing in lymphonodes, appears to be particularly useful for the cellular and the in-vivo microscopy image processing in which few a priori assumption on the type, the extent and the variability of particle motions, can be done.
authors.affiliation: Dipartimento di Fisica, Università degli Studi di Milano Bicocca, Milano, Italy.
authors.forename: Carlo E
authors.initials: CE
authors.lastname: Villa
authors.forename: Michele
authors.initials: M
authors.lastname: Caccia
authors.forename: Laura
authors.initials: L
authors.lastname: Sironi
authors.forename: Laura
authors.initials: L
authors.lastname: D'Alfonso
authors.forename: Maddalena
authors.initials: M
authors.lastname: Collini
authors.forename: Ilaria
authors.initials: I
authors.lastname: Rivolta
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Miserocchi
authors.forename: Tatiana
authors.initials: T
authors.lastname: Gorletta
authors.forename: Ivan
authors.initials: I
authors.lastname: Zanoni
authors.forename: Francesca
authors.initials: F
authors.lastname: Granucci
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Chirico
mesh_terms: D000465:Algorithms; D000818:Animals; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D042541:Intracellular Space / Q000378:metabolism; D008214:Lymphocytes / Q000166:cytology*; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D013997:Time Factors
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