Microparticle sizing by dynamic light scattering in fresh-frozen plasma

• Published in: Vox sanguinis Vol. 96; no. 3; pp. 206 - 212
• Main Authors: Lawrie, A. S., Albanyan, A., Cardigan, R. A., Mackie, I. J., Harrison, P.
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01-04-2009; S. Karger AG
• Subjects: Blood banks; Blood products; Cell-Derived Microparticles - chemistry; DLS; dynamic light scattering; FFP; Freezing; Humans; Light; microparticles; Particle Size; Plasma; Plasma - chemistry; Scattering, Radiation; Scientific method
• ISSN: 0042-9007; 1423-0410
• DOI: 10.1111/j.1423-0410.2008.01151.x

Background   We have previously shown that fresh‐frozen plasma (FFP) contains red blood cell‐derived procoagulant microparticles (MPs) that are removable by 0·2 µm filtration. Given the limitations of current methods for accurately sizing MPs, we have applied the novel approach of dynamic light scattering (DLS) to characterize the size distributions of these MPs within FFP.
Methods   Fresh‐frozen plasma was prepared from blood Group A and O donations (n = 10 of each) after an overnight hold of whole blood at 4°C. On the day of analysis, plasma was thawed to 37°C and daughter aliquots were studied pre‐ and post‐filtration (0·2 µm filtration device, Ceveron® MFU‐500, Technoclone). MP size and dispersity was assessed using a Zetasizer Nano S (Malvern Instruments Ltd), which employs a 173° backscatter detector and an N5 Submicron Particle Size Analyser (Beckman Coulter) using multi‐angle measurements (30·1°, 62·6° and 90°). The analysers presented MP size distribution graphically as intensity plots, mean size, standard deviation and polydispersity index.
Results   Of the instruments used, only the N5 utilizing a 30·1° angle of measurement could detect MPs of the expected size distribution and demonstrate their removal by filtration. MPs (range of mean particle diameters: pre, 101–464 nm; post, 21–182 nm filtration) were significantly smaller post‐filtration (P < 0·0001), but polydispersity index (median: pre, 0·746, post, 0·769) exhibited no significant change. There was no significant difference between the size of MPs from blood Group O (pre, 247 nm) and Group A (pre, 289 nm) samples (P = 0·44).
Conclusion   Our data demonstrates that DLS offers a novel approach to assessing MP size and distribution, a technique that could be easily adopted as a means of assessing MPs within either FFP or other blood products.