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Measurement of Particle Size Distribution in Turbid Solutions by Dynamic Light Scattering Microscopy
Published on: January 9, 2017
Microparticle sizing by dynamic light scattering in fresh-frozen plasma
A S Lawrie1, A Albanyan, R A Cardigan
1Haemostasis Research Unit, Department of Haematology, University College London, 51 Chenies Mews, London, UK. andrew.lawrie@ucl.ac.uk
Background:
We have previously shown that fresh-frozen plasma (FFP) contains red blood cell-derived procoagulant microparticles (MPs) that are removable by 0.2 microm 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 degrees C. On the day of analysis, plasma was thawed to 37 degrees C and daughter aliquots were studied pre- and post-filtration (0.2 microm filtration device, Ceveron MFU-500, Technoclone). MP size and dispersity was assessed using a Zetasizer Nano S (Malvern Instruments Ltd), which employs a 173 degrees backscatter detector and an N5 Submicron Particle Size Analyser (Beckman Coulter) using multi-angle measurements (30.1 degrees , 62.6 degrees and 90 degrees ). 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 degrees 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.
Insights
Dynamic Light Scattering (DLS) accurately sizes red blood cell microparticles (MPs) in fresh-frozen plasma (FFP). Filtration significantly reduces MP size, confirming DLS as a valuable tool for blood product quality control.
Area of Science:
- Hematology
- Biotechnology
- Analytical Chemistry
Background:
- Fresh-frozen plasma (FFP) contains procoagulant microparticles (MPs) derived from red blood cells.
- Current methods for sizing MPs have limitations.
- Dynamic Light Scattering (DLS) is a novel approach for MP characterization.
Purpose of the Study:
- To characterize the size distribution of MPs in FFP using DLS.
- To evaluate the effectiveness of 0.2 micrometer filtration in removing MPs.
- To assess the suitability of DLS for quality control of blood products.
Main Methods:
- FFP from blood groups A and O was analyzed using DLS (Zetasizer Nano S and N5 Submicron Particle Size Analyser).
- MP size and dispersity were measured pre- and post-0.2 micrometer filtration.
- Multi-angle measurements were employed to assess MP size distribution.
Main Results:
- The N5 analyser at a 30.1-degree angle detected MPs and confirmed their removal by filtration.
- Mean MP diameter significantly decreased post-filtration (101-464 nm pre vs. 21-182 nm post).
- No significant change in polydispersity index was observed; MP size did not differ significantly between blood groups O and A.
Conclusions:
- DLS is a novel and effective method for assessing MP size and distribution in FFP.
- The findings support the use of DLS for quality assessment of FFP and other blood products.
- 0.2 micrometer filtration effectively reduces the concentration of procoagulant MPs in FFP.
