Abstract
Microsample analysis is highly beneficial in blood-based testing where cutting-edge bioanalytical technologies enable the analysis of volumes down to a few tens of microliters. Despite the availability of analytical methods, the difficulty in obtaining high-quality and standardized microsamples at the point of collection remains a major limitation of the process. Here, we detail and model a blood separation principle which exploits discrete viscosity differences caused by blood particle sedimentation in a laminar flow. Based on this phenomenon, we developed a portable capillary-driven microfluidic device that separates blood microsamples collected from finger-pricks and delivers 2 µL of metered serum for bench-top analysis. Flow cytometric analysis demonstrated the high purity of generated microsamples. Proteomic and metabolomic analyses of the microsamples of 283 proteins and 1351 metabolite features was consistent with samples generated via a conventional centrifugation method. These results were confirmed by a clinical study scrutinising 8 blood markers in obese patients.
Highlights
Blood sample separation, consisting in the extraction and isolation of the liquid surrounding blood cells, is the most common preparation operation performed before clinical biochemistry analysis
In this paper we report a device developed for sample preparation at the point of collection (SP-POC) that addresses the need for a blood microsample separation device that generates standardized and stabilized
The duration of the separation delay is dependent on cell sedimentation speed and distance: the process is influenced by blood parameters and design parameters but is independent of whole blood feeding flowrate
Summary
Blood sample separation, consisting in the extraction and isolation of the liquid surrounding blood cells, is the most common preparation operation performed before clinical biochemistry analysis. Most common microsystems for blood separation traditionally rely on sedimentation[2,3], microfiltration[4,5] or cell deviation[6,7] in a microfluidic chip[8] (see Supplementary Note 1). Typically, such systems require either sample pre-dilution, have complex designs or suffer from low extraction yields[9]. Mass spectrometry proteomics and metabolomics profiling, as well as standard clinical chemistry analysis, were performed on the chip-separated samples to demonstrate their quality and their suitability for gold standard bench-top analysis methods
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