Single particle tracking

Principle of single particle tracking: The rectangles represent frames from an image acquisition at times t=0,1,2,.... The tracked particles are represented as red circles and in the last frame, the reconstructed trajectories are shown as blue lines

Single particle tracking (SPT) is the observation of the motion of individual particles within a medium. The coordinates (x, y, z) over a series of time steps is referred to as a trajectory. The trajectory can be analyzed to identify modes of motion or heterogeneities in the motion such as obstacles or regions of fast transport (e.g. due to active transport or flow), in the case of random motion trajectory analysis can provide a diffusion coefficient.

Applications

1. Single Particle Tracking is used to quantify specific behaviour of a protein on the cell surface. 2. It can also use to understand the cellular kinetics of many proteins like HIV-1 Tat Protein Transduction domain (Tat-PTD) [1]

Brownian diffusion

Main article: Brownian motion

Membrane diffusion

Components of biological membranes diffuse in a viscous pseudo-two-dimensional environment. By labeling membrane component with either an optical label (such as a gold or polystyrene bead) or a fluorescent tag a trajectory can be observed. The trajectory of lipids and proteins within the membrane of live cells have been used to infer the mechanisms affecting their motion.[2] One of the methods to observe optical labels on cell membranes is known as nanovid microscopy. Another interesting technique associated with particle tracking is real-time 3D particle tracking, that was used in the context of observing small complexes as they are shuttled around in or near the cell membrane.[3]

Methods

Particle tracking can be performed using fluorescent or optical labels. The use of optical labels, such as gold nanoparticles, in SPT experiments has been termed nanovid microscopy. In addition, there are several imaging modalities for 3D particle tracking including Multifocal plane microscopy,[4] and DH-PSF microscopy.[5]

See also

References

  1. ↑ Single Particle Tracking Confirms That Multivalent Tat Protein Transduction Domain-induced Heparan Sulfate Proteoglycan Cross-linkage Activates Rac1 for Internalization-Junji Imamura, Yasuhiro Suzuki, Kohsuke Gonda, Chandra Nath Roy, Hiroyuki Gatanaga, Noriaki Ohuchi, Hideo Higuchi-Journal of Biological Chemistry, Vol. 286, No. 12. (25 March 2011), pp. 10581-10592. doi:10.1074/jbc.M110.187450
  2. ↑ 1. Saxton, M. J., Jacobson, K. Single-particle tracking: Applications to membrane dynamics. Annual Review of Biophysics and Biomolecular Structure 1997;26(373-399).
  3. ↑ 2. Hellriegel, C., Gratton, E. Real-time multi-parameter spectroscopy and localization in three-dimensional single-particle tracking. J R Soc Interface (2009) 6:S3-14.
  4. ↑ Ram, Sripad; Prabhat, Prashant; Chao, Jerry; Sally Ward, E.; Ober, Raimund J. (2008). "High accuracy 3D quantum dot tracking with multifocal plane microscopy for the study of fast intracellular dynamics in live cells". Biophysical Journal 95 (12): 6025–6043. doi:10.1529/biophysj.108.140392.
  5. ↑ Badieirostami, M.; Lew, M.D.; Thompson, M.A.; Moerner, W.E. (2010). "Three-dimensional localization precision of the double-helix point spread function versus astigmatism and biplane". Applied Physics Letters 97: 161103. doi:10.1063/1.3499652.