TY - JOUR
T1 - Intracellular Dynamics of Extracellular Vesicles by Segmented Trajectory Analysis
AU - Rautaniemi, Kaisa
AU - John, Thomas
AU - Richter, Maximilian
AU - Huck, Benedikt C.
AU - Zini, Jacopo
AU - Loretz, Brigitta
AU - Lehr, Claus Michael
AU - Vuorimaa-Laukkanen, Elina
AU - Lisitsyna, Ekaterina
AU - Laaksonen, Timo
N1 - Funding Information:
The macrorheological characterization of HEC was done by S. M. Recktenwald, Saarland University. This work was supported by the Academy of Finland under Grant 311362 (K.R., E.L., E.V.-L.), 316893 (T.L.), 314406 (J.Z.), and 323669 (E.L.); Business Finland EVE ecosystem under Grant 1842/31/2019 (K.R., J.Z., E.L., E.V.-L.); Alfred Kordelin Foundation (K.R.); BMBF project ANTI-TB fund # GWANTA20 (B.C.H.); and ERC CoG, Grant agreement No 101001016 (T.L.).
Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.
PY - 2022/12/27
Y1 - 2022/12/27
N2 - The analysis of nanoparticle (NP) dynamics in live cell studies by video tracking provides detailed information on their interactions and trafficking in the cells. Although the video analysis is not yet routinely used in NP studies, the equipment suitable for the experiments is already available in most laboratories. Here, we compare trajectory patterns, diffusion coefficients, and particle velocities of NPs in A549 cells with a rather simple experimental setup consisting of a fluorescence microscope and openly available trajectory analysis software. The studied NPs include commercial fluorescent polymeric particles and two subpopulations of PC-3 cell-derived extracellular vesicles (EVs). As bioderived natural nanoparticles, the fluorescence intensities of the EVs limited the recording speed. Therefore, we studied the effect of the recording frame rate and analysis parameters to the trajectory results with bright fluorescent commercial NPs. We show that the trajectory classification and the apparent particle velocities are affected by the recording frame rate, while the diffusion constants stay comparable. The NP trajectory patterns were similar for all NP types and resembled intracellular vesicular transport. Interestingly, the EV movements were faster than the commercial NPs, which contrasts with their physical sizes and may indicate a greater role of the motor proteins in their intracellular transports.
AB - The analysis of nanoparticle (NP) dynamics in live cell studies by video tracking provides detailed information on their interactions and trafficking in the cells. Although the video analysis is not yet routinely used in NP studies, the equipment suitable for the experiments is already available in most laboratories. Here, we compare trajectory patterns, diffusion coefficients, and particle velocities of NPs in A549 cells with a rather simple experimental setup consisting of a fluorescence microscope and openly available trajectory analysis software. The studied NPs include commercial fluorescent polymeric particles and two subpopulations of PC-3 cell-derived extracellular vesicles (EVs). As bioderived natural nanoparticles, the fluorescence intensities of the EVs limited the recording speed. Therefore, we studied the effect of the recording frame rate and analysis parameters to the trajectory results with bright fluorescent commercial NPs. We show that the trajectory classification and the apparent particle velocities are affected by the recording frame rate, while the diffusion constants stay comparable. The NP trajectory patterns were similar for all NP types and resembled intracellular vesicular transport. Interestingly, the EV movements were faster than the commercial NPs, which contrasts with their physical sizes and may indicate a greater role of the motor proteins in their intracellular transports.
U2 - 10.1021/acs.analchem.2c02928
DO - 10.1021/acs.analchem.2c02928
M3 - Article
AN - SCOPUS:85143965721
SN - 0003-2700
VL - 94
SP - 17770
EP - 17778
JO - Analytical Chemistry
JF - Analytical Chemistry
IS - 51
ER -