Fig 1.
(A) Schematic representation of protocol used for the isolation of CSF microvesicles and exosomes. (B) In nanoparticles tracking analysis, light scattered by EVs is captured by digital camera over a series of frames. The rate of the particle movement is then used to calculate particle size using the Stokes—Einstein equation. (C) In tunable resistive pulse sensing, EVs change the electrical resistance as they pass through a pore-based sensor resulting in a resistive pulse signal. Signals obtained from the measurements can then be used to calculate the size, concentration and charge of each particle by correlating the signal back to a set of known standards. (D) In Vesicle flow cytometry, EVs were stained with an optimized concentration of a fluorogenic lipophilic probe, di-8-ANEPPS, and detected on a custom high sensitivity flow cytometer. Vesicle diameter was estimated by comparison to di-8-stained liposomes.
Fig 2.
Comparison of EV quantification by NTA and TRPS.
EVs were isolated from CSF collected from glioblastoma patients by differential centrifugation into microvesicle (10,000×g) and exosome (120,000×g) fractions, and resuspended in PBS. Isolated EVs were analyzed by NTA or TRPS. (A) Size profile of CSF exosomes determined by NTA and TRPS. (B) Size profile of CSF microvesicles determined by NTA and TRPS. (C) Comparison of EV yield by size ranges.
Table 1.
Total EV yield as determined by NTA and TRPS.
Table 2.
EV yield by size range as determined by NTA and TRPS.
Fig 3.
Comparison of EV quantification by NTA and VFC.
CSF EVs isolated by differential centrifugation into microvesicle (10,000×g) and exosome (120,000×g) fractions were analyzed by NTA or VFC. (A) Size profile of CSF exosomes determined by NTA and VFC. (B) Size profile of CSF microvesicles determined by NTA and VFC. (C) Comparison of EV yield by size ranges.
Table 3.
Total EV as determined by NTA and VFC.
Table 4.
EV yield by size range as determined by NTA and VFC.
Fig 4.
Comparison of EV quantification by NTA and TEM.
CSF EVs were fractionated into microvesicles (10,000×g) and exosomes (120,000×g) by differential ultracentrifugation and then analyzed by NTA and TEM. (A) Representative TEM images, scale bar = 200nm. (B) Total EV count as determined by NTA and TEM. Fold difference in particle detected between NTA and TEM is denoted.
Table 5.
Total EV as determined by NTA and TEM.