Accumulative Difference image protocol for particle tracking in fluorescence microscopy tested in mouse lymphonodes

Carlo E. Villa, Michele Caccia, Laura Sironi, Laura D'Alfonso, Maddalena Collini, Ilaria Rivolta, Giuseppe Miserocchi, Tatiana Gorletta, Ivan Zanoni, Francesca Granucci, Giuseppe Chirico

Research output: Contribution to journalArticle

3 Citations (Scopus)

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.

Original languageEnglish
Article numbere12216
JournalPLoS One
Volume5
Issue number8
DOIs
Publication statusPublished - 2010

Fingerprint

Fluorescence microscopy
fluorescence microscopy
Fluorescence Microscopy
Cytology
Lymphocytes
mice
Edge detection
Image segmentation
microscopy
Microscopic examination
Image processing
Fluorescence
image analysis
Nanoparticles
Color
medical sciences
Lipids
Imaging techniques
Cell Biology
Noise

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Accumulative Difference image protocol for particle tracking in fluorescence microscopy tested in mouse lymphonodes. / Villa, Carlo E.; Caccia, Michele; Sironi, Laura; D'Alfonso, Laura; Collini, Maddalena; Rivolta, Ilaria; Miserocchi, Giuseppe; Gorletta, Tatiana; Zanoni, Ivan; Granucci, Francesca; Chirico, Giuseppe.

In: PLoS One, Vol. 5, No. 8, e12216, 2010.

Research output: Contribution to journalArticle

Villa, CE, Caccia, M, Sironi, L, D'Alfonso, L, Collini, M, Rivolta, I, Miserocchi, G, Gorletta, T, Zanoni, I, Granucci, F & Chirico, G 2010, 'Accumulative Difference image protocol for particle tracking in fluorescence microscopy tested in mouse lymphonodes', PLoS One, vol. 5, no. 8, e12216. https://doi.org/10.1371/journal.pone.0012216
Villa, Carlo E. ; Caccia, Michele ; Sironi, Laura ; D'Alfonso, Laura ; Collini, Maddalena ; Rivolta, Ilaria ; Miserocchi, Giuseppe ; Gorletta, Tatiana ; Zanoni, Ivan ; Granucci, Francesca ; Chirico, Giuseppe. / Accumulative Difference image protocol for particle tracking in fluorescence microscopy tested in mouse lymphonodes. In: PLoS One. 2010 ; Vol. 5, No. 8.
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